Isivf09 Brochure Final

SPONSOR ACKNOWLEDGEMENT
We thank the following companies for their generous support of the Congress by providing
unrestricted educational grants:
ELITE SPONSOR
Merck Serono
PREMIER SPONSOR
IBSA
CORPORATE SPONSOR
Schering-Plough
SUPPORTER
Ferring Pharmaceuticals
FINAL PROGRAMME & ABSTRACTS
TABLE OF CONTENTS
General Information
ISIVF Membership Secretariat 2 Committees
2155 Guy Street, 14th Floor
Room 1471
3 Message from the President of ISIVF
Montreal, Quebec, Canada H3H 2R9 3 About ISIVF
Tel.: (514) 843-1556 4 Message from the President of ISIVM
Fax: (514) 843-2880
info@isivf.com
5 Message from the Congress President
6 CME Information
7 Congress Information
i 8 Map / Floor Plans
Siv 10
12
Local Information
Social Programme & Accompanying Persons Tour
Scientific Programme
2009 Congress Secretariat 15 Agenda-at-a-Glance
c/o IS Event Solutions 16 Instructions to Oral and Poster Presenters
1334 Notre-Dame West, 2nd Floor
Montréal, Québec, Canada H3C 1K7 17 ISIVF/ISIVM Faculty
Tel.: (514) 392-7703 19 Industry-Sponsored Symposia
Fax: (514) 227-5083 21 Sunday – April 19 Pre-Congress Courses
info@iseventsolutions.com
22 Monday – April 20 Programme
29 Tuesday – April 21 Programme
Printed in Switzerland, April 2009 35 Wednesday – April 22 Programme
Abstracts
41 Monday
51 Tuesday
65 Wednesday
73 Posters
99 Exhibit Directory
100 Exhibit Floor Plan
101 Exhibit Hours
101 List of Exhibitors
102 Exhibitor Details
107 Presenter Index
110 Certificate of Attendance
Geneva
1
15th World Congress on In Vitro Fertilization | 4th World Congress on In Vitro Maturation
COMMITTEES
Organizing Committee L. MENCAGLIA
JEAN-BERNARD DUBUISSON, Chair Department of Obstetrics and Gynecology,
Geneva University Hospitals, Geneva
BARRY CAPPEL
M. MUELLER
ANIS FEKI Universitaetsklinik fuer Frauenheilkunde, Inselsspital Bern
VICTOR GOMEL P. PETIGNAT
SEANG LIN TAN Department of Obstetrics and Gynecology,
Scientific Programme Committee P.A. SAPPINO
Service d'oncologie, Geneva University Hospitals, Geneva
JEAN-BERNARD DUBUISSON
D. STUCKI
ANIS FEKI Clinique de gynécologie et d'obstétrique,
VICTOR GOMEL Hopital Cantonal de Fribourg, Fribourg
TIMUR GURGAN S. VAN DER POEL

World Health Organization, Geneva
SEANG LIN TAN
International Scientific Committee
Local Scientific Committee
MICHEL ABOU ABDALLAH DAVID GARDNER
SE. ANTONARAKIS Beirut, Lebanon Melbourne, Australia
Department of Genetic Medicine and Development, University of Geneva
Medical School, and University Hospitals of Geneva, Geneva MOHAMED ABOULGHAR VICTOR GOMEL
Cairo, Egypt Vancouver, Canada
P. BISCHOF
Department of Obstetrics and Gynecology, AYDIN ARICI TIMUR GURGAN
Geneva University Hospitals, Geneva New Haven, United States Ankara, Turkey
C. DE GEYTER PHILIPPE BOUCHARD MILTON LEONG

Gynecologic Endocrinology, Universitätsspital Basel, Basel Paris, France Hong Kong, China
KEITH CAMPBELL RAGAA MANSOUR
J.B. DUBUISSON - Department of Obstetrics and Gynecology, Loughborough, UK Cairo, Egypt
RI-CHENG CHIAN GIANPIERO PALERMO
P. FEHR Montreal, Canada New York, United States
Euro Haus, Schaffhausen
KLAUS DIEDRICH ZEV ROZENWAKS
A. FEKI Luebeck, Germany New York, United States
Department of Obstetrics and Gynecology,
Geneva University Hospitals, Geneva JACQUES DONNEZ HASSAN SALLAM
Brussels, Belgium Smouha, Egypt
M. GERMOND
Centre de Procréation Médicalement Assistée et d'Endocrinologie JEAN-BERNARD DUBUISSON BILL SCHOOLCRAFT
Geneva, Switzerland Englewood, United States
Gynécologique (CPMA), Lausanne
ANIS FEKI LYNETTE SCOTT
M. HOHL Geneva, Switzerland Middleton, United States
Frauenklinik, Kantonsspital Baden, Baden
RENÉ FRYDMAN SEANG LIN TAN
O. HOVATTA Clamart, France Montreal, Canada
Department of Clinical Science, Intervention and Technology, Division of
Obstetrics and Gynecology, Karolinska Institutet, Stockholm, Sweden JUAN A. GARCIA VELASCO
Madrid, Spain
B. IMTHURN
Department of Obstetrics and Gynaecology,
University Hospital Zurich, Zurich
M. JACONI Scientific Programme Contributers
Department of Pathology and Immunology,
Faculty of Medicine, Geneva University, Geneva ALPHA Scientists in Reproductive Medicine
K.-H. KRAUSE
APART
Département d'Immunologie et Pathologie, Middle Eastern Fertility Society
Faculté de médecine de Genève, Geneva Turkish Society of Reproductive Medicine
April 19-22, 2009 2

WELCOME MESSAGE
WELCOME FROM THE PRESIDENT OF ISIVF
Dear Colleagues, Ladies and Gentlemen,
It is my particular pleasure and honor to welcome you to the 15th World Congress on IVF,
jointly held with the 4th World Congress on IVM here in Geneva. First of all, I would like to
express my cordial gratitude to the Congress President Prof. Dubuisson, the members of the
scientific program committee and the organizing committee for their elaborate and dedicated
contribution, which is the key to providing a quality programme, leading to overall success of the Congress. Looking at
the marvelously constructed program, I believe that the Congress shall enjoy a great deal of success, giving satisfaction
to every participant.
Since the birth of the first IVF baby achieved by Professor Robert Edwards, our Honorary Patron, IVF-related science
and technology has widened and deepened during the past 30 years in an unexpectedly speedy pace to establish a
systematized infertility treatment discipline called assisted reproductive technology (ART). World IVF Congress, founded
by the late Professor Kurt Semm in 1980, has consistently served as the central venue of forum responsible for the
reproductive revolution. With these changes occurring in ART worldwide, it was proposed and agreed at the Board
Meeting of the 13th World Congress in Istanbul to establish the International Society for IVF (ISIVF), for innovation in
its structure and function as a scientific body, to cover all the disciplines and address concerns of all professionals
associated with ART.
I wish you a wonderful Congress and a memorable stay in Geneva!
Takahide Mori
President, International Society for In Vitro Fertilization
ABOUT ISIVF
ISIVF's primary mission is to promote research and clinical development of in vitro fertilization of human oocytes (IVF)
for the treatment of infertility and as an Assisted Reproduction Technology. Objectives include:
1 Promulgating ethical practice and standards of practice of IVF treatment;
2 Fostering collaboration between the various centres in the world that offer IVF treatment;
3 Organizing symposia for the purposes of presentation, discussion, exchange and publication of
research, data, knowledge, theories and standards pertaining to IVF,
and specifically, a forum under the name “World Congress on In Vitro Fertilization".
3 Geneva
WELCOME MESSAGE
WELCOME FROM THE PRESIDENT OF ISIVM
Dear colleagues,
On behalf of the Organizing and Scientific Committees it

is my pleasure to welcome you to the 4th World
Congress on In Vitro Maturation. We are delighted to be
holding this year’s congress in the beautiful city of In conclusion, I would like to thank the Congress
Geneva. President, Professor Jean-Bernard Dubuisson; Professor
Takahide Mori, President of the International Society for
It is especially fitting that Geneva serves as the European In Vitro Fertilization; and the members of the Scientific
headquarters for the United Nations, a global and Organizing Committees for all of their hard work in
organization that promotes international cooperation and making this meeting possible. My sincere thanks are
understanding. As speakers and delegates from around also extended to the speakers and session chairs for
the world assemble for this year’s congress, the parallel their participation in this year’s program.
nature of our missions becomes very apparent. We are
here from many nations to share the latest advancements It is our sincere hope that you will enjoy the Congress
and to provide new insights into finding the best and all of the charm that Geneva has to offer. We hope
possible ways to treat the growing problem of infertility that you will take home with you new and valuable
– a problem that affects all cultures and nationalities. information about IVM that will assist you in your future
practice and research.
Because it does not require the use of fertility drugs, IVM
has become an increasingly important method of
treatment and its applications are widening to include
fertility preservation, particularly for patients with cancer,
autoimmune disorders, severe endometriosis or Turner
Mosiac. This year’s scientific program has been
designed to provide you with a comprehensive overview Seang Lin Tan
and understanding of this procedure as well as to President, International Society for In Vitro Maturation
address some of the concerns related to it.
However, the learning process is ongoing and keeping

up with the latest advances and their attendant
controversies is a demanding task. The ISIVM Society
was formed to foster the continued exchange of ideas
and knowledge and to promote the ethical, standardized
practice of IVM. If you have not already done so, I urge
you to join this important organization to share in its
many benefits. Please visit our website at
www.isivm.com for further information.
April 19-22, 2009 4

WELCOME MESSAGE
WELCOME FROM THE CONGRESS PRESIDENT
Dear colleagues,
Geneva is an international business and art centre. It is

the meeting point of world diplomacy; its political,
economic and cultural aura combined with an excellent interaction between the protagonists in these different
tourist trade. All this, make Geneva a major metropolis of fields. To further enhance this interaction and dialogue,
world fame. Great international organizations like the Red the International Board of ISIVF 2009 took the initiative to
Cross, the International Labour Office, the World Health drive forward this international Congress in Geneva.
Organization, have their headquarters here. It is of
importance to mention that Geneva has a long tradition of Reproduction and stem cell biology are by any measure
watch making (Baume et Mercier, Charriol, Chopard, considered amongst the currently most dynamic research
Franck Muller, Patek Philippe, Rolex, Raymond Weil, fields in life science. This International Congress will
Omega, etc.). span several of these areas. Important topics are to review
the current advances in the treatment of infertility couples,
With this small introduction I wish to welcome you to the including controlled ovarian stimulation, natural cycles,
city of Geneva for the 15th World Congress on In Vitro endometriosis, low cost IVF and fertility preservation.
Fertilization, and the 4th World Congress on In Vitro This Congress will cover the current status of stem cell
Maturation. research for potential therapy for the field of reproduction.
The Geneva ISIVF Congress will bring together leading Welcome to Geneva, the city of Calvin. I am sure that you
scientists and clinicians from various areas of infertility, will enjoy these few days in Geneva, the “Capital of
ART, IVM and stem cell research. Peace”, the International city of new ideas and
The inspiration to organise such conference came from humanitarian laws!
the importance and the necessity of close and continuous
Jean-Bernard Dubuisson
Congress President
5 Geneva
CME INFORMATION
CME Credits Learning Objectives
This event is approved for up to 27 credits by the
Centre for Continuing Medical Education (“CME”). The General goals for event:
Centre for CME, Faculty of Medicine, McGill University is
fully accredited by the Committee on Accreditation of As a result of participation and interaction in this
Canadian Medical Schools and through the (CACMS) is congress, participants will be able to:
accredited to award AMA PRA category 1 credits. • Expand their broad understanding of the current
state of the art in In Vitro Fertilization and In Vitro
This event is an Accredited Group Learning Activity as Maturation
defined by the Maintenance of Certification programme of • Gain an in-depth knowledge of selected focused
the Royal College of Physicians and Surgeons of Canada. subject matter at the leading edge of current
The Centre for CME, Faculty of Medicine, McGill developments in the fields of IVM and IVF
University designates this activity for Category 1 credit • Interact and network with key individuals in the
towards the AMA Physicians Recognition Award up to the field
maximum number of credit hours noted above. Each
physician should claim only those hours of credit that • Know about research being done at present that
he/she actually spent at the educational activity. will impact on their practice in the future
• Become re-energized in their professional lives by
Delegates requiring CME Credits must sign in at a sense of shared experiences and collegiality
the Congress Services Desk every day that they
are present at the congress and must collect Knowledge participants gain:
their Attestation CME Certificate on the last day
of their attendance at the congress. • Review of all IVF, IVM and ART related topics (all-
inclusive programme)
Congress Evaluation • Update on most recent advances
A Congress evaluation form is included in your delegate • Close look at latest stem cell research
bag. Please complete it and drop it off at the Congress • Research content and clinical science will be
Evaluation mailbox located in the Registration Area.The discussed
Committee appreciates your constructive feedback.
CONGRESS INFORMATION
Best Abstract Awards
(provided by the International Society for In Vitro Fertilization)
Two abstract awards winners, one for poster and one for
oral presentation, will be selected. The winners will be
announced in the Closing Ceremonies.
Certificate of Attendance
The certificate of attendance is included as last page of
this publication. You can simply remove the certificate
and fill in your name.
April 19-22, 2009 6

CONGRESS INFORMATION
Lost and Found
Delegate Hospitality Any lost and found items will be held at the
Congress Services desk for the duration of the event.
• Badge Cord For any unclaimed items after the congress, please
Graciously sponsored by Schering Plough contact IS Event Solutions.
• Delegate Bag Luggage

Graciously sponsored by Merck Serono We recommend that you leave your luggage with the
Bellman at your hotel. The Convention Centre does
• Notepad and Pen not have a bellman or concierge service. The
Graciously sponsored by IBSA Pharmaceuticals organizers don't take any responsibility for suitcases
that are left anywhere at the Convention Centre,
• Coffee Breaks including the registration desk.
Graciously sponsored by Ferring Pharmaceuticals
Daily coffee breaks will be served in the exhibit area. For Poster Area
exact timing, please refer to the detailed programme. The poster area is available at all times towards the
back of the exhibit area. Delegates can visit posters
• Delicious and Affordable Lunches are served at the at their convenience. Poster presenters are
cafeteria on Level 1. Refreshments and snacks can be encouraged to be at their posters during break times.
purchased at the Café.
Registration Information
Emergency / First Aid
For any emergency or first aid services inside the Convention • Registration Hours
Centre, please dial “9112” from any phone. Sunday, April 19 15:00 - 20:00
Monday, April 20 07:30 - 18:00
Information Tuesday, April 21 07:30 - 18:00
Should you require any assistance during the Congress, Wednesday, April 22 07:30 - 18:00
please call the Congress Services desk directly at +41 22
791 94 92. The Conference Services Desk is located in the • Name Badges
registration area and is in service for the entire duration of the For security purposes, all congress participants
Congress. must wear their name badges when onsite at the
CICG.
Internet Access
OPTION 1: Internet Stations • Badge Colours
The CICG provides 4 Internet stations which are located in the Magenta Delegates and sponsors
exhibit area. These stations allow for Internet access to check Blue Invited speakers
your emails via a web browser. You can not print, upload or Red Executive Board members
download files from these stations. The terminals will operate Green Exhibitors
during exhibit hours. Orange Single day registrants
Yellow Guests
OPTION 2: Wireless Internet (wi-fi) Black Media representatives
Wireless Internet is available for delegates carrying a laptop White Staff
and can be accessed anywhere inside the CICG. Laptops must
be wi-fi enabled. Public Notice
The Convention Centre (CICG) is a smoke-free
Username: isivf Password: isivf environment. Smoking is permitted only outside the
building.
7 Geneva
Coordonnées
Centre International de
Conférence
t
Genève (CICG)
17 rue de Varembé
CH 1211 Genève 20
Tél. +41 (0)22 791 91 11
Fax +41 (0)22 791 90 64
Centre de Conférences
de Varembé
t
(CCV)
9-11 rue de Varembé
CH 1211 Genève 20
Tél. +41 (0)22 791 91 11
Fax +41 (0)22 791 90 64
Entrée
>
Principale
>
Entrée
Espace Dunand
Les accès
Access
Adresses utiles
Genève Tourisme
Aéroport 18 rue du Mt-Blanc - CP 1602
international CH 1211 Genève 20
Tél. +41 (0)22 909 70 00
Fax +41 (0)22 909 70 01
Genève
Aéroport International
CP 100 - CH 1215 Genève 15
Tél. +41 (0)22 717 71 11
Fax +41 (0)22 798 43 77
CCV Gare de Cornavin

CH 1200 Genève
CICG Tél. 0900 300 300
Lac de Genève
ou
Le Léman
Gare
Cornavin
April 19-22, 2009 8

L
FLOOR PLANS
Location of Rooms 2-3-4 (Level 0)
Location of Room 18 (Level -1)
9 Geneva
LOCAL INFORMATION
About Geneva Currency

Some call it the "world's smallest metropolis", others the The Swiss franc (CHF). 1 CHF divided into 100
"peace capital". Whatever image it brings to mind, Geneva centimes is the national currency. Automatic teller
has always been a popular destination, known throughout the machines and exchange offices are readily available.
world as the United Nations' European headquarters and the Most hotels, restaurants and shops accept major
head office of the International Committee of the Red Cross. credit cards.
Geneva is the smallest of the world's major cities. It is a world
apart, with something for everyone. Distinguished by its At press time, the following exchange rates apply
unique geographical position in the heart of Europe, state-of- (April 2009).
the-art technology, high-quality services, prestige and ranking
1 USD 1.14 CHF
as a world-class city, coupled with the advantages of a small
1 EURO 1.54 CHF
town, Geneva is the ideal venue for our World Congress.
Geneva is magnificently situated on the banks of the largest 100 JAPAN YEN 1.18 CHF
lake in central Europe, at the foot of the Jura mountains. 1 CHINA YUAN RENMINBI 0.66 CHF
At the gates of the Alps, Geneva has all that is needed
Getting Around Geneva
to charm you.
Unireso is the tariff community covering all of
Geneva.Tickets and day passes must be purchased
The town, whose rich past still comes to life today, is the
before boarding the vehicle from automatic vending
true international capital and can offer visitors a great number
machines located at the stops. Ticketing machines
of different aspects. These include its humanitarian
accept Swiss and Euro coins. A ticket costs CHF 3.00
commitment, its varied cultural activities, its major
and a 1-day pass costs CHF 10. Most hotels provide
congresses and exhibitions and its renowned cuisine,
a complimentary transportation card for all of Geneva.
beautiful countryside and opportunities for many different
excursions. The beauty of the site, the close proximity, a
Important Telephone Numbers
prestigious Alpine region and its privileged location along the
When calling from within Switzerland, you always
main axes of the West, make Geneva naturally one of the
have to dial the area code 22 with a “0” in front of it,
largest European centres of tourism.
even if you're in the same area. To call from within
Geneva or other places in Switzerland, dial 022 + area
Communication
code + local number.
Public telephones are easy to locate and international calling
cards may be used. They can be purchased at most
To dial a number overseas, press 00 + country code +
convenience stores and newsstands. Major credit cards are
area code + telephone number.
also acceptable billing alternatives.
Hotel Information
Credit Cards
InterContinental Geneva (0) 22 919 39 39
In order to report lost or stolen credit cards, these numbers
Hotel Le Royal (0) 22 906 14 14
will provide assistance.
Hotel Auteuil (0) 22 544 22 22
Hotel Epsom (0) 22 544 66 66
American Express +41 44 659 63 33
Hotel Kipling (0) 22 544 40 40
Mastercard 0 800 89 4732
Visa 0 800 89 7092
Other Important Numbers
Taxi Service (0) 22 33 141 33
Tourism Geneva (0) 22 909 70 00
April 19-22, 2009 10

LOCAL INFORMATION
Electricity Taxes and Tax Refund

The current used throughout Switzerland is 230 Volts The VAT you pay on purchased goods in Switzerland is
(AC), 50 cycles. Most power sockets are designed for 7.6%. You may ask at the shops for your Global Refund
three pin round plugs. The standard continental type plug Cheque and reclaim the VAT. The total purchases in a shop
with two round pins, applied for many electrical travel must amount to CHF 400.00 (including VAT). The tourist
products, may be used without problems. Adaptors are must be resident outside Switzerland and the goods must be
available in most hotels. exported within 30 days.
Languages 3 easy steps to claiming your refund in Switzerland

The official language of the Conference will be English. IN THE STORE:
The language most frequently spoken in Geneva is Your total purchases in a shop must amount to CHF 400.00
French. However, you should have no problem (Including VAT). You must be a resident outside Switzerland
conversing in English. and the goods must be exported within 30 days.
Shopping THROUGH CUSTOMS:

Geneva is a shopper's paradise! With a myriad of When leaving Switzerland the Tax-free Shopping Cheques
boutiques and department stores, Geneva offers have to be stamped by Swiss customs authorities after they
something for everyone. It's the watch capital of the have seen the goods.
world, a center for exquisite jewelry, and a place to find
high quality Swiss and imported items. Before you leave, COLLECTING THE REFUND:
don't forget to purchase some chocolate from one of You have several choices: immediate cash at a Cash
Geneva's master chocolate-makers. And why not stock up Refund Office, direct crediting to a chosen credit card or
on Swiss army knives? They make the ideal bank account, a bank check and even, for certain countries,
gift for anyone - including yourself! a cash refund when you return home.
Store Hours:
Monday - Wednesday 9 am-7 pm
Thursday 9 am-9 pm
Friday 9 am-7.30 pm
Saturday 9 am-6 pm
Sunday closed
Time Difference
GMT +1 hour
11 Geneva
SOCIAL PROGRAMME &

ACCOMPANYING PERSONS TOUR
Official Opening Ceremonies & Welcome Reception
Sunday, April 19, 17:00 to 20:00
Centre de conférence international de Genève (CICG)
(included in full registration fee for delegates and registered accompanying persons)
Dress Code: Casual

After the initial inauguration of the Congress with light entertainment, delegates will have a chance to meet with
colleagues and exhibitor representatives as we inaugurate the congress exhibit with a welcome reception, where light
cocktails and refreshments will be served.
Accompanying Persons Tour

Tuesday, April 21, 10:00-17:00
Starting point: Centre de conférence international de Genève (CICG)
The first part of the tour is a city tour which takes you to the International part of Geneva, through the centre of the
world's peacemaking organizations followed by a tour of the main tourist attractions of the city. Get a taste of Geneva's
history and dig even deeper into the Old Town on a guided walking tour. The day will then continue with a countryside
excursion which will take you around Lake Geneva and through charming vineyards, castles and wonderful
fortified houses.
April 19-22, 2009 12

SCIENTIFIC PROGRAMME
13 Geneva
NOTES
April 19-22, 2009 14

AGENDA AT A GLANCE
15 Geneva
INSTRUCTIONS FOR ORAL

AND POSTER PRESENTERS
Speaker Preparation Hours
The speaker check-in is located in front of room 2.
All speakers must visit the speaker check-in in order to upload the presentation and sign required paperwork.
SUNDAY, APRIL 19 14:00-20:00

MONDAY TO WEDNESDAY, APRIL 20-22 07:30-18:00
Speakers MUST check in and submit their PowerPoint presentation at the Speaker Check-in outside room 2 at least
24 hours before their scheduled presentation time and revisions may be made up to 3 hours prior to presentation.
This is the only way that we can be sure that you are present, and that the Chairs of your session are updated on any
last-minute changes. It is also very important that you ensure the seamless functioning of your data presentation.
A technician will load your presention onto the main network server. Your presentation will be on the network and
therefore available when you arrive in your session room.
Please note that individual laptop computers are not permitted for presentations.
Poster Presenters
Participants will view posters during lunches and breaks, therefore authors are encouraged to be present at their
posters during those times.
Authors are responsible for the setting up and the removal of their posters according to the following schedule:
Mounting time: MONDAY, APRIL 20 07:00-08:30

Removal: WEDNESDAY, APRIL 22 BY 18:00
Posters not removed by the specified time on the last day of their presentation will be removed and discarded by
Meeting staff. ISIVF cannot accept liability for lost or damaged posters. ISIVF will not mail posters to authors after
the meeting.
April 19-22, 2009 16

ISIVF/ISIVM FACULTY
A b i r, R o n i t Fa dini, Rube ns
Rabin Medical Center, Petah Tikva, Israel Biogenesi Reproductive Medicine Center, Monza, Italy
Abou Abda l la h, Mic he l Fe k i, Anis
MEFS, Beirut, Lebanon Geneva University Hospitals, Geneva, Switzerland
A h u j a , K a ma l Fe nic he l, Pa t ric k
The London Women’s Clinic, London, UK Centre Hospitalo-Universitaire de Nice, Nice, France
A l b e r t i n i , Da v i d F r y d m a n , Ne l l y
Kansas Life Science Innovations Center, Kansas City, USA Hôpital Antoine Béclère, Clamart, France
Ambrose t t i, Ale xa ndra Fr ydma n, Re né
Geneva University Hospitals, Geneva, Switzerland Hôpital Antoine Béclère, Clamart, France
B e n -Yo s e f , Da l i t F u l k a J r., J o s e f
The Tel Aviv Sourasky Medical Center, Tel Aviv, Israel Institute of Animal Science, Prague, Czech Republic
Be nk ha lif a , Monc e f Ge r m o n d , M a r c
ATL R&D, Reproductive Biology and Genetics, La Verrière, France Centre de Procréation Médicalement Assistée et d’Endocrinologie
Gynécologique (CPMA), Lausanne, Switzerland
Bisc hof , P a ul
Geneva University Hospitals, Geneva, Switzerland Gi l b e r t , Lu c y
McGill University, Royal Victoria Hospital, Montreal, Canada
Boivin, Ja c k y
Cardiff University, School of Psychology, Cardiff, UK Go m e l , V i c t o r
University of British Columbia, Vancouver, Canada
Bouc ha r d, Philippe
Hôpital Saint Antoine, Paris, France Go u g e o n , A l a i n
Centre hospitalier Lyon-Sud, Lyon, France
Buc k e t t , Willia m
McGill University, Royal Victoria Hospital, Montreal, Canada Gü r g a n , T i m u r
Gürgan Clinic, Ankara, Turkey
Ca m p b e l l , K e i t h
University of Nottingham - School of Biosciences, Loughborough, Ha n c k , B e v e r l y
UK Infertility Awareness Association of Canada, Montreal, Canada
Ch i a n , R i -Ch e n g Ha n d y s i d e , A l a n
McGill University, Royal Victoria Hospital, Montreal, Canada The London Bridge Fertility Gynaecology and Genetic Centre,
London, UK
Ch i l l i k , Cl a u d i o
Matercell, Buenos Aires, Argentina Ho l z e r, Ha n a n e l
McGill Reproductive Centre, Montreal, Canada
Da l Ca n t o , M a r i a b e a t r i c e
Istituti Clinici Zucchi, Monza, Italy Ho v a t t a , Ou t i
Karolinska Institutet, Stockholm, Sweden
d e Ge y t e r, Ch r i s t i a n
University of Basel, Basel, Switzerland Ishiz uk a , Bunpe i
Saint Marianna University School of Medicine, Kanagawa, Japan
De m i r o l , Ay g ü l
Gürgan Clinic, Ankara, Turkey J o n e s , K e i t h T.
The University of Newcastle Australia, Newcastle, Australia
Di e d r i c h , K l a u s
Universitätsklinikum Schleswig-Holstein, Luebeck, Germany J o s s o , Na t h a l i e
INSERM, Clamart, France
Do n n e z , J a c q u e s
Saint-Luc University, Brussels, Belgium K a t o , Os a m u
Kato Ladies Clinic, Shinjuku-ku, Tokyo, Japan
Do r, J e h o s h u a
The Joseph Buchman Gynecology and Maternity Center, Le d g e r, W i l l i a m
Sheba Medical Center, Israel Jessop Hospital for Women, Sheffield, UK
Du b u i s s o n , J e a n -B e r n a r d Le o n g , M i l t o n
Geneva University Hospitals, Geneva, Switzerland Hong Kong Sanatorium and Hospital, Hong Kong, China
17 Geneva
ISIVF/ISIVM FACULTY
Le s s e y , B r u c e Se r m o n , K a r e n
Center for Women’s Medicine, Greenville, USA Uze Universitat Brussel, Brussels, Belgium
Le u n g , P e t e r C.K . Sh o h a m , Ze e v
The University of British Columbia, Vancouver, Canada Kaplan Hospital, Rehovot, Israel
Li m , J i n Ho Si l b e r, Sh e r m a n
Maria Fertility Hospital, Seoul, Korea The Infertility Center of St. Louis, Saint Louis, USA
Li n d e n b e r g , Sv e n d Ta k e f m a n , J a n e t
Nordica Fertility Clinic, Institut for Human Reproduktion, McGill Reproductive Centre, Montreal, Canada
Copenhagen, Denmark
Ta n , Se a n g Li n
M a n s o u r, R a g a a McGill Reproductive Centre, Montreal, Canada
The Egyptian IVF-ET Center, Cairo, Egypt
Tu r-K a s p a , I l a n
M e n c a g l i a , Lu c a Reproductive Genetics Institute, Chicago, USA
Florence Centre of Ambulatory Surgery & Infertility, Florence, Italy
Ur m a n , B u l e n t
M e t t l e r, Li s e l o t t e American Hospital of Istanbul, Nisantasi, Istanbul, Turkey
University of Kiel, Kiel, Germany
v a n d e r P o e l , Sh e r y l
M i k k e l s e n , A n n Li s UNDP/UNFPA/WHO/World Bank, Geneva, Switzerland
Holbæk Hospital, Holbæk, Denmark
v a n He r e n d a e l , B r u n o J .
M o r i m o t o , Yo s h i h a r u Endoscopic Training Centre Antwerp (ETCA), Antwerp, Belgium
IVF Namba Clinic, Osaka, Japan
V a ss e n a , R i t a
M u e l l e r, M i c h a e l D. Centre for Regenerative Medicine, Barcelona, Spain
Bern University Hospital, Bern, Switzerland
V e r h a a k , Ch r i s
Murdoc h, A lison Radboud Universtiy Medical Center, Nijmegen, The Netherlands
International Centre for Life, Newcastle upon Tyne, UK
Wa t r e l o t , A n t o i n e
Na r d o , Lu c i a n o Centre de Recherche et d’Etude de la Stérilité, Lyon, France
St. Mary’s Hospital, Manchester, UK
We n g e r, J e a n -M a r i e
P a l e r m o , Gi a n p i e r o Geneva University Hospitals, Geneva, Switzerland
Weill Cornell Medicine College, New York, USA
Wo o d r u f f , Te r e s a K .
Pe t igna t , Pa t ric k Northwestern University Feinberg School of Medicine, Evanston, USA
Geneva University Hospitals, Geneva, Switzerland
Yo u n g , Lo r r a i n e
P o u l y, J . Lu c University of Nottingham, Nottingham, UK
Polyclinique de l’Hotel Dieu, Clermont-Ferrand, France
Za l e l , Ya r o n
P r a s a d , Su d h a Sheba Medical Center, Tel Hashomer, Israel
Maulana Azad Medical College, New Delhi, India
Zo r n , B r a n k o
Qi a o , J i e University Medical Center Ljubljana, Ljubljana, Slovenia
The Third School of Clinical Medicine, Beijing, China
R a i n e -F e n n i n g , Ni c h o l a s
University of Nottingham, Nottingham, UK
R e m o h í Gi m e n e z , J o s é
University of Valencia & Equipo IVI, Valencia, Spain
Sa l l a m , Ha s s a n
University of Alexandria and Alexandria Fertility Center,
Alexandria, Egypt
Sc h e n k e r, J o s e p h
Madassah Medical Center Hebrew University, Jerusalem, Israel
April 19-22, 2009 18

INDUSTRY-SPONSORED SYMPOSIA
Merck Serono Satellite Symposium GONADOTROPINS FUNCTIONAL PROJECT I:

Monday, April 20 | 12:45-14:00 UNDERSTANDING THE MOLECULAR BASIS OF
Room 2 GONADOTROPIN ACTION IN THE GONADAL AXIS
AND THE CLINICAL IMPLICATIONS IN rFSH/rLH
PROTOCOLS OF STIMULATION FOR ART
CHAIR: Marc Germond (Switzerland)
SPEAKERS:
MOLECULAR ACTION OF LH ON THE FOLLICLE

Claus Yding Andersen (Denmark)
THE ROLE OF rFSH/rLH IN FOLLICULAR DEVELOPMENT

AND THE GENETICS OF THE LH RECEPTOR
Dimitris Loutradis (Greece)
CLINICAL EXPERIENCE WITH A NEW COMBINATION OF

RFSH AND RLH IN A 2:1 RATIO FOR ART: SPANISH
PRELIMINARY RESULTS
Jose Luis Caballero (Spain)
IBSA Satellite Symposium HELPING YOU TO IMPROVE YOUR SUCCESS

Tuesday, April 21 | 12:45-14:00 RATE
Room 2
CHAIRMAN: B. Imthurn (Switzerland)
SPEAKERS:
DOES THE hCG DOSE FOR FINAL FOLLICULAR

MATURATION HAVE AN IMPACT ON THE IVF
OUTCOMES?
Christian de Geyter (Switzerland)
ARE THESE DIFFERENT HMGS?

Carlo Alviggi (Italy)
IVF IN THE REAL WORLD: SOCIAL AND ETHICAL

CONCERNS
Gillian Lockwood (UK)
19 Geneva
NOTES
April 19-22, 2009 20

PRE-CONGRESS COURSES
SUNDAY, APRIL 19, 2009 | HOTEL INTERCONTINENTAL GENEVA
08:30-12:00 Morning Courses
Course 1: Endometriosis and Infertility Room: Madrid

100.1 Jacques Donnez, Brussels, Belgium
100.2 Jean-Bernard Dubuisson, Geneva, Switzerland
100.3 Victor Gomel, Vancouver, Canada
100.4 Michael Mueller, Bern, Switzerland
100.5 Jean-Marie Wenger, Geneva, Switzerland
Course 2: PGD and ART Outcomes Room: Rome

101.1 René Frydman, Clamart, France
101.2 Alan Handyside, London, UK
101.3 Nelly Frydman, Clamart, France
101.4 Karen Sermon, Brussels, Belgium
Course 3: Stem Cells and Reprogramming Room: Brussels

102.1 Outi Hovatta, Stockholm, Sweden
102.2 Anna Veiga, Barcelona, Spain
102.3 Anis Feki, Geneva, Switzerland
12:00-13:00 Lunch Break
13:00-16:30 Afternoon Courses
Course 4: Ultrasonography and ART Room: Madrid

103.1 Nicholas Raine-Fenning, Nottingham, UK
103.2 Ilan Tur-Kaspa, Chicago, USA
103.3 Yuron Zalel, Tel Hashomer, Israel
Course 5: Germ Cell and Tissue Ovarian

Preservation: Impact on IVM Development Room: Rome
104.1 Seang Lin Tan, Montreal, Canada
104.2 Timur Gurgan, Ankara, Turkey
104.3 Moncef Benkhalifa, La Verriere, France
Course 6: Low Cost IVF Room: Brussels

105.1 Sheryl van der Poel, WHO, Geneva, Switzerland
WHO and International Initiatives
105.2 Lisa Feldman
WHO Primary Health Care Management of the Infertile Couple
105.3 Dr. Meserat Mengistu, Addis Ababa and Umeå, Sweden
The Need and Present Situation in Women's Health and Infertility
Treatment in Ethiopia
105.4 Outi Hovatta, Karolinska Institutet, Stockholm, Sweden
Experience from a Low Cost IVF Unit in Khartoum, Sudan
21 Geneva
DETAILED PROGRAMME
MONDAY, APRIL 20, 2009
08:30-10:15 Plenary I Room: Salle 2

Co-Chairs: Gomel, Victor - Vancouver, Canada
Palermo, Gianpiero - New York, USA
200.1 ALPHA KEYNOTE LECTURE - PGD: CURRENT DEVELOPMENT AND FUTURE DIRECTIONS
Handyside, Alan - London, UK
200.2 NATURAL CYCLE IVF/IVM
Lim, Jin Ho - Seoul, Korea
200.3 ONCOFERTILITY: THE PRESERVATION OF FERTILITY OPTIONS FOR YOUNG
PEOPLE WITH CANCER
Woodruff, Teresa K. - Evanston, USA
10:15-10:45 Coffee Break, Exhibits and Poster Viewing
10:45-12:30 Concurrent Symposium 1 - Implantation Room: Salle 3
Co-Chairs: Bischof, Paul - Geneva, Switzerland

Chian, Ri-Cheng - Montreal, Canada
Prasad, Sudha - New Delhi, India
201.1 LUTHEAL SUPPORT IN IVF: WHY, WITH WHAT, WHEN TO START AND FOR HOW LONG?
Shoham, Zeev - Rehovot, Israel
201.2 ASSESSMENT AND REGULATION OF ENDOMETRIAL RECEPTIVITY
Lessey, Bruce - Greenville, USA
201.3 CELLULAR AND MOLECULAR MECHANISMS ALLOWING EMBRYO IMPLANTATION
Bischof, Paul - Geneva, Switzerland
201.4 ENDOMETRIAL IMAGING AND ART OUTCOME
Tur-Kaspa, Ilan - Chicago, USA
10:45-12:30 Concurrent Symposium 2 - Ultrasound and Controlled Room: Salle 2

Ovarian Stimulation
Co-Chairs: Raine-Fenning, Nicholas - Nottingham, UK

202.1 TRANSVAGINAL HYDROLAPAROSCOPY IN INFERTILITY
Watrelot, Antoine - Lyon, France
202.2 3DUS AND UTERINE PATHOLOGIES
Zalel, Yaron - Tel Hashomer, Israel
202.3 OVARIAN IMAGING & ART OUTCOME
Raine-Fenning, Nicholas - Nottingham, UK
202.4 ULTRASOUND UTERINE INVESTIGATION IN ART
Qiao, Jie - Beijing, China
April 19-22, 2009 22

DETAILED PROGRAMME
10:45-12:30 ALPHA Session (Scientists in Reproductive Medicine) Room: Salle 4
Co-Chairs: Handyside, Alan - London, UK

Balaban, Basak - Istanbul, Turkey
203.1 EPIGENETICS AND SAFETY IN ART

Young, Lorraine - Nottingham, UK
203.2 EFFECT OF OVARIAN STIMULATION ON EMBRYO CHROMOSOMES
203.3 PREGNANCY AND BIRTH OUTCOME FOLLOWING PGD
12:30-14:00 Lunch Break, Exhibits and Poster Viewing
12:45-14:00 Merck Serono Satellite Symposium (refer to page 19 for details)
14:00-15:45 Concurrent Symposium 3 - Adnexal Surgery and Fertility Room: Salle 3
Co-Chairs: Lebbi, Issam - Tunisia

Mueller, Michael D. - Bern, Switzerland
Dubuisson, Jean-Bernard - Geneva, Switzerland
204.1 REPRODUCTIVE LIFE AFTER GYNECOLOGICAL CANCER
Petignat, Patrick - Geneva, Switzerland
204.2 IS RESECTION OF ENDOMETRIOMAS PRIOR TO IVF/ICSI NECESSARY?
Gürgan, Timur - Ankara, Turkey
204.3 OVARIAN ADHESIOLYSIS AND FERTILITY
Nardo, Luciano - Manchester, UK
204.4 OVARIAN FUNCTION AND REGULATION
Mettler, Liselotte - Kiel, Germany
204.5 HYSTEROSCOPY IN INFERTILITY AND IVF/ICSI TREATMENT
Demirol, Aygül - Ankara, Turkey
23 Geneva
DETAILED PROGRAMME
14:00-15:45 Concurrent Symposium 4 - Premature Ovarian Failure Room: Salle 2
Chair: Tavmergen, Ege - Izmir, Turkey

205.1 ETIOLOGICAL FACTORS AND CLINICAL COURSE IN PATIENTS WITH PREMATURE OVARIAN FAILURE
Ishizuka, Bunpei - Kanagawa, Japan
205.2 OVARIAN FAILURE AND FERTILITY
Bouchard, Philippe - Paris, France
205.3 REGULATION OF GRANULOSA CELL APOPTOSIS BY GNRH
Leung, Peter C.K. - Vancouver, Canada
205.4 G-CSF AS NON-INVASIVE APPROACH FOR OOCYTE COMPETENCE
Frydman, René - Clamart, France
14:00-15:45 Swiss Society of Reproductive Medicine Session - (SSRM) Room: Salle 4

Research Prizes of SSRM: The Best Papers
Co-Chairs: Imthurn, Bruno - Zurich, Switzerland

Galié-Wunder, Dorothea - Lausanne, Switzerland
206.1 HIGH SURVIVAL AND DEVELOPMENTAL RATES OF VITRIFIED MOUSE ZYGOTES FOLLOWING
POLAR BODY BIOPSY
Macas, M. - Zurich, Switzerland
206.2 COMPUTER-ASSISTED ANALYSIS OF HUMAN PRONUCLEAR ZYGOTES
Urner, F. - Lausanne, Switzerland
206.3 MATURE OVARIAN FOLLICLES CONTAIN A SUBPOPULATION OF STEM CELLS
Kossowska-Tomaszczuk, K. - Basel, Switzerland
206.4 INTERMEDIATE AND PREMUTATION FMR1 ALLELES IN WOMEN WITH OCCULT PRIMARY
OVARIAN INSUFFICIENCY
Streuli, I. - Geneva, Switzerland
206.5 GENE EXPRESSION PROFILING OF CULTURED ENDOMETRIUM FROM WOMEN UNDERGOING
IN VITRO FERTILIZATION WITH DIFFERENT OUTCOME
Bersinger, N. - Bern, Switzerland
206.6 TASK FORCE OF THE FERTILITY AND CANCER NETWORK IN THE FRENCH SPEAKING PART OF
SWITZERLAND (RÉSEAU ROMAND CANCER ET FERTILITÉ)
Ambrosetti, Alexandra - Geneva Switzerland
April 19-22, 2009 24

DETAILED PROGRAMME
14:00-15:45 Oral Communications 1 Room: Salle 18
Co-Chairs: Petignat, Patrick - Geneva, Switzerland

Gurgan, Timur - Ankara, Turkey
207.1 DETECTION OF OXIDATIVE DNA DAMAGE BY FLOW CYTOMETRY
AND ITS ASSOCIATION WITH MALE INFERTILITY
Nozha, Chakroun Feki - Sfax, Tunisia
207.2 IMPRINTING IN THE HUMAN EMBRYO: EVIDENCE FOR A RECYCLING OF HOMOCYSTEINE IN THE OOCYTE
Benkhalifa, Moncef - La Verrière, France
207.3 HIV SERO-CONVERSION OF A SPERM DONOR DURING THE DONATION PERIOD
Smith, Venessa – London, UK
207.4 PRESENTATION OF INNOVATIVE PATENTED PRODUCTS FOR OVUM PICK UP AND EMBRYO TRANSFER
Steiner, Hans-Peter - Graz, Austria
207.5 COMBINING TWO ZWITTERIONIC BUFFERS IN IVF HANDLING MEDIA SUPPORTS MOUSE BLASTOCYST
DEVELOPMENT AND NORMAL HUMAN OOCYTE FERTILIZATION FOLLOWING ICSI
Swain, Jason - Ann Arbor, USA
207.6 CORRELATION BETWEEN REACTIVE OXYGEN SPECIES (ROS) IN SEMINAL PLASMA AND SPERM VITALITY,
MEMBRANE, DNA INTEGRITY, DNA STRAND BREAKS OF INFERTILE MEN BEFORE AND AFTER SEMEN
PROCESSING BY PURESPERM AND IVF/ICSI OUTCOMES
Hammadeh, M.E. - Homburg/Saar, Germany
16:15-18:00 Concurrent Symposium 5 - Development of IVM Room: Salle 2
Co-Chairs: Tan, Seang Lin - Montreal, Canada

Mikkelsen, Ann Lis - Holbæk, Denmark
208.1 NUCLEOLI IN MAMMALIAN OOCYTES AND EARLY EMBRYOS
Fulka Jr., Josef - Prague, Czech Republic
208.2 EARLY OVULATION INDUCTION: IS IT A METHOD TO REPLACE IVM AND PREVENT OHSS
Pouly, J. Luc - Clermont-Ferrand, France
208.3 AN OOCENTRIC VIEW OF FOLLICULOGENESIS AND EMBRYOGENESIS
Albertini, David - Kansas City, USA
208.4 IVM FOR REGULAR MENSTRUAL CYCLING WOMEN
Fadini, Rubens - Monza, Italy
208.5 CURRENT CRITICISM ON IVM TREATMENT
Leong, Milton - Hong Kong, China
25 Geneva
DETAILED PROGRAMME
16:15-18:00 Concurrent Symposium 6 - Laboratory and Management Room: Salle 3
Chair: Gülekli, Bülent - Balcova Izmir, Turkey

209.1 EMBRYO SELECTION
Hovatta, Outi - Stockholm, Sweden
209.2 INTRODUCING ISO INTO IVF ACADEMIC UNITS
Frydman, Nelly - Clamart, France
209.3 ISO STANDARDS FOR CLINICAL WORKUP - EXPERIENCES WITH DIN EN ISO 9001:2000
Imthurn, Bruno - Zurich, Switzerland
209.4 OBSTETRIC AND NEONATAL OUTCOME FOLLOWING IVM
Buckett, William - Montreal, Canada
16:15-18:00 Turkish Society of Reproductive Medicine Session Room: Salle 4
Co-Chairs: Gürgan, Timur - Ankara, Turkey

Pabuçcu, Recai - Turkey
210.1 GENETIC APROCHES UNVEIL CHECKPOINTS DURING FOLLICULAR MATURATION
Benkhalifa, Moncef
210.2 NOVEL TECHNOLOGIES IN ART LABORATORIES: FACTS OR ARTEFACTS?
Fındıklı, Necati
210.3 WHAT TO DO AFTER ART FAILS
Göker, Ege
210.4 OFFICE HYSTEROSCOPY PRIOR ICSI/IVF : DOES IT MAKE ANY DIFFERENCE?
Demirol, Aygül
210.5 SURGERY BEFORE IVF/ICSI : CAN WE IMPROVE THE SUCCESS RATES
Tavmergen, Erol
210.6 COMPARISON OF EFFICACY OF REC-FSH AND HIGHLY PURIFIED HUMAN-DERIVED FSH IN GNRH
ANTAGONIST CYCLES FOR ART
Bahçeci, M.
210.7 COMPARISON OF EFFICACY OF REC-FSH AND HIGHLY PURIFIED HUMAN-DERIVED FSH IN GNRH
ANTAGONIST CYCLES FOR ART
Ulug, U.
April 19-22, 2009 26

DETAILED PROGRAMME
Chair: Holzer, Hananel - Montreal, Canada

211.1 EFFECT OF SEASON ON FERTILIZATION AND EMBRYO QUALITY RATES IN ART
Danfour, Mohamed - Misurata, Libya
211.2 THE ROLE OF IMMUNOHYSTOCHEMISTRY IN COMPLEX MORPHOLOGICAL
DIAGNOSIS OF WOMEN STERILITY
Kostyuchek, Irina - Saint-Petersburg, Russia
211.3 WHAT WE OFFER TO WOMEN BEFORE UNDERGOING SURGICAL TREATMENT FOR MENORRHAGIA
Younas, Kinza - Birmingham, UK
211.4 GLYCODELIN ENHANCES FERTILIZATION DURING IVF
Gneist, Nadja - Dresden, Germany
211.5 A SYSTEMATIC REVIEW OF MANAGEMENT OF INFERTILITY IN WOMEN OVER THE AGE OF 40
Marinakis, Gerasimos - London, UK
211.6 ROLE OF DIAGNOSTIC LAPAROSCOPY IN WOMEN WITH NORMAL PELVIC IMAGING AND FAILED SUPER
OVULATION AND INTRA UTERINE INSEMINATION
Jayakrishnan, Kay - Trivandrum, India
211.7 EFFECTIVE WAY TO CUT THE POST OPERATIVE ANALGESIC REQUIREMENT AFTER FEMALE TUBAL
STERILISATION
Younas, Kinza - Birmingham, UK
27 Geneva
NOTES
April 19-22, 2009 28

DETAILED PROGRAMME
TUESDAY, APRIL 21, 2009
08:30-10:15 Plenary II Room: Salle 2
Co-Chairs: Dubuisson, Jean-Bernard - Geneva, Switzerland

Campbell, Keith - Loughborough, UK
300.1 ENDOMETRIOSIS RELATED INFERTILITY: THE NEED FOR A CONSERVATIVE SURGERY
Donnez, Jacques - Brussels, Belgium
300.2 NEW CHALLENGES FOR OVARIAN-CELL THERAPY AT THE DAWN OF THE 21ST CENTURY
Gougeon, Alain - Lyon, France
300.3 IVM AND FERTILITY PRESERVATION
Tan, Seang Lin - Montreal, Canada
10:45-12:30 Concurrent Symposium 7 - Controlled Ovarian Stimulation Room: Salle 4
Co-Chairs: Kato, Osamu - Tokyo, Japan

Tavmergen, Erol - Izmir, Turkey
Bouchard, Philippe - Paris, France
301.1 RECURRENT IMPLANTATION FAILURE: A CLINICIAN'S VIEW
Schenker, Joseph - Jerusalem, Israel
301.2 PHARMACOLOGICAL REGIMENTS FOR UTERINE PREPARATION FOR ET
301.3 MULTIPLE PREGNANCY, IS IT A FAILURE OF ART?
Belaisch-Allart, Joelle - Sèvres, France
301.4 DIAGNOSTIC AND MANAGEMENT OF POOR RESPONDERS
Germond, Marc - Lausanne, Switzerland
301.5 LUTEAL SUPPORT IN ART
Prasad, Sudha - New Delhi, India
10:45-12:30 Concurrent Symposium 8 - Fertility Preservation Room: Salle 3
Co-Chairs: Rosenwaks, Zev - New York, USA

302.1 CONSEQUENCES OF CHEMOTHERAPY AND/OR RADIOTHERAPY ON FERTILITY
Dor, Jehoshua - Tel Aviv, Israel
302.2 CRYOPRESERVATION OF OOCYTES
302.3 VITRIFICATION OF HUMAN ZYGOTES: CLINICAL RESULTS
Diedrich, Klaus - Luebeck, Germany
29 Geneva
DETAILED PROGRAMME
10:45-11:25 APART Symposium 1 - Natural Cycle IVF and Mild IVF Cycle Room: Salle 2
in Reproductive Medecine
Co-Chairs: Feichtinger, Wilfried - Vienna, Australia

Kamiya, Hirofumi - Sapporo, Japan
303.1 THE ROLE OF MILD STIMULATION IN THE CURRENT IVF PRACTICE
Bernard, Artur - Budapest, Hungary
303.2 OUTCOME OF THE RECENT IVF PROGRAM TRANSFERRING SINGLE BLASTOCYST RETRIEVED ON
MINIMAL STIMULATION CYCLE AND FROZEN BY VITRIFICATION METHOD
Kato, Keiichi - Tokyo, Japan
303.3 CONTROLLED OVARIAN STIMULATION, MINIMAL STIMULATION OR NO STIMULATION FOR IN VITRO
FERTILIZATION: IS THERE AN OPTIMAL STIMULATION?
Zhang, John - New York, USA
11:25-12:30 APART Symposium 2 - Management of PCO Patients Room: Salle 2
Co-Chairs: Barak, Yona - Berzliya-On-Sea, Israel

Morimoto, Yoshiharu - Sapporo, Japan
304.1 THE BENEFIT OF IVM IN PCO PATIENTS
Barak, Yona - Berzliya-On-Sea, Israel
304.2 OXYGEN CONSUMPTION BY SCANNING ELECTROCHEMICAL MICROSCOPY (SECM) FOR HUMAN IVM COC
& EMBRYO WITH PCO PATIENTS
Yoshida, Hiroaki - Sendai, Japan
304.3 CLINICAL DATA OF IVM FOR PCO PATIENTS
Morimoto, Yoshiharu - Osaka, Japan
304.4 USE OF LETROZOLE IN PCOS
Shozu, Makio - Chiba, Japan
304.5 PCOS: MANAGEMENT STRATEGIES IN 2009
Yelian, Frank - Los Angeles, USA
12:45-14:00 IBSA Satellite Symposium (for details, please refer to page 19)
April 19-22, 2009 30

DETAILED PROGRAMME
14:00-15:45 Concurrent Symposium 9 - Fertility Preservation Room: Salle 4
Co-Chairs: Donnez, Jacques - Brussels, Belgium

Diedrich, Klaus - Luebeck, Germany
Ishizuka, Bunpei - Kanagawa, Japan
305.1 MANAGEMENT OF OVARIAN TUMORS IN CONTEXT OF FERTLITY PRESERVATION
Gilbert, Lucy - Montreal, Canada
305.2 FRESH AND FROZEN OVARY TRANSPLANTATION
Silber, Sherman - Saint Louis, USA
305.3 PRESERVING FERTILITY IN CANCER PATIENTS AND FOLLICLE OVARIAN RESERVE
Donnez, Jacques - Brussels, Belgium
305.4 OOCYTE VITRIFICATION FOR CANCER AND NON-CANCER FERTILITY PRESERVATION
Tan, Seang Lin - Montreal, Canada
14:00-15:45 Concurrent Symposium 10 - IVM Room: Salle 3
Chair: Leong, Milton - Hong Kong, China

306.1 REGULATION OF SPINDLE AND CHROMATIN DYNAMICS DURING OOCYTE MATURATION
Smith, Gary - Ann Arbor, USA
306.2 FOLLICULAR DEVELOPMENT IN VIVO AND IN VITRO
Abir, Ronit - Petah Tikva, Israel
306.3 ULTRASTRUCTURE OF IN VITRO MATURED OOCYTES
Morimoto, Yoshiharu - Osaka, Japan
306.4 GENOMIC IMPRINTING IN ART
14:00-14:50 APART Symposium 3 - ART for Fertility Preservation Room: Salle 2
Co-Chairs: Kaali, Steven - New York, USA

Utsunomiya, Takafumi - Oita, Japan
307.1 FERTILITY PRESERVING OPTIONS OF REPRODUCTIVE AGE CANCER PATIENTS
Kovacs, Peter - Budapest, Hungary
307.2 THE CURRENT APPROACH TO OOCYTE VITRIFICATION FOR CANCER PATIENTS IN JAPAN
Utsunomiya, Takafumi - Oita, Japan
307.3 SUCCESSFUL VITRIFICATION OF HUMAN OOCYTES
Kuwayama, Masashige - Tokyo, Japan
307.4 CRYOPRESERVATION OF PREPUBERTAL TESTICULAR TISSUE
Yoshida, Atsumi - Tokyo, Japan
31 Geneva
DETAILED PROGRAMME
14:50-15:45 APART Symposium 4 - Male Infertility Room: Salle 2
Co-Chairs: Zech, Herbert - Bregenz, Austria

Tanaka, Atsushi - Fukuoka, Japan
308.1 DOES A COMBINATION OF CHINESE HERBS AND ACUPUNCTURE TREATMENT AFFECT SPERM
CHARACTERISTICS IN INFERTILE COUPLE
Velleman, Ramon - Tel Aviv, Israel
308.2 IMPROVEMENT OF A PHOTODYNAMIC SYSTEM FOR OBSERVATION OF SEMINIFEROUS TUBULES
IN MICRODISSECTION-TESTICULAR SPERM EXTRACTION (MD-TESE)
Tanaka, Atsushi - Fukuoka, Japan
308.3 A NOVEL CRYOPRESERVATION TECHNIQUE FOR VERY FEW MOTILE SPERM FROM SEVERELY
INFERTILE MEN
Kyono, Koichi - Sendai, Japan
308.4 ARGUMENTS TO IMPLEMENT THE SELECTION OF SPERMATOZOA AT HIGH MAGNIFICATION BEFORE ICSI
Zech, Herbert - Bregenz, Austria
Chair: Germond, Marc - Lausanne, Switzerland

309.1 SEARCH FOR MECHANISMS UNDERLYING IMPRINTING DEFECTS IN SPERM
Naumova, Anna K. - Montreal, Canada
309.2 EFFECT OF MATURATION IN VITRO ON SPINDLE MORPHOLOGY IN HUMAN OOCYTES
Munk, Mette - Copenhagen, Denmark
309.3 PATERNAL AGE AFFECTS SPERM DNA FRAGMENTATION BUT NOT ITS PACKAGING
Belloc, Stéphanie - Paris, France
309.4 THE LOCALIZATION OF SPECIFIC PROTEIN KINASE C ISOTYPES IN MOUSE OVARY
Tepekoy, Filiz - Antalya, Turkey
309.5 IMPACT OF A GNRH ANTAGONIST ON THE OUTCOME OF CONTROLLED OVARIAN
HYPERSTIMULATION (COH) IN WOMEN WITH POLYCYSTIC OVARY SYNDROME:
A PROSPECTIVE AND RANDOMIZED STUDY
Stadtmauer, Laurel - Norfolk, USA
309.6 OVARIAN AGING: PROGNOSTIC FACTORS, BIOLOGICAL FACTORS AND OUTCOME OF ART
Genazzani, Andrea Riccardo - Pisa, Italy
309.7 FIRST TIME EVIDENCE FOR INCREASED PLACENTAL GROWTH FACTOR MRNA IN ENDOMETRIUM OF
PATIENTS WITH SUCCESSFUL IMPLANTATION
Santi, Alessandro - Bern, Switzerland
April 19-22, 2009 32

DETAILED PROGRAMME
Co-Chairs: Tan, Seang Lin - Montreal, Canada

Albertini, David - Kansas City, USA
310.1 EFFICIENCY OF IVM EGG COLLECTION
Gülekli, Bülent - Izmir, Turkey
310.2 FSH PRIMING FOR IVM
Mikkelsen, Ann Lis - Holbæk, Denmark
310.3 HCG PRIMING FOR IVM
Son, Weon-Young - Montreal, Canada
310.4 ENDOMETRIAL PREPARATION IN IVM TREATMENT
Lindenberg, Svend - Copenhagen, Denmark
16:15-18:00 Concurrent Symposium 12 - PGD Room: Salle 2
Co-Chairs: Frydman, Nelly - Clamart, France

Antonarakis, Stylianos - Geneva, Switzerland
311.1 PRE IMPLANTATION GENETIC DIAGNOSIS (PGD) FOR CANCER PREDISPOSITION GENES
Yaron, Yuval - Tel Aviv, Israel
311.2 PGD IMPROVED PGD TESTS AND CLINICAL APPLICATIONS
Frydman, René - Clamart, France
311.3 PGD VS PRE-GENETIC SCREENING (PGS): WHERE IS THE LIMIT?
Gianaroli, Luca - Bologna, Italy
33 Geneva
DETAILED PROGRAMME
Co-Chairs: Benkhalifa, Moncef – La Verrière, France

Gougeon, Alain - Lyon, France
312.1 ULTRASOUND GUIDED VERSUS BLIND TOUCH EMBRYO TRANSFER: THE EGYPTIAN EXPERIENCE
Taha, Tamer Fouad - Cairo, Egypt
312.2 A MICROSYSTEM WITH WHITE LIGHT OPTICAL SENSOR TO QUALIFY THE CYTOPLASM MATURITY
OF HUMAN OOCYTES
Roux, Christophe - Besançon, France
312.3 HYSTEROSCOPIC ENDOMETRIAL EMBRYO DELIVERY (HEED): TRANSFER OF DAY 2-5 EMBRYOS
Kamrava, Michael - Beverly Hills, USA
312.4 NOVEL METHOD OF HAND FREE ULTRASOUND QUIDED ET
Haloob, A. Rahim - Basildon, UK
312.5 CERVICAL MUCUS STATUS CAN BE ACCURATELY ESTIMATED BY TRANSVAGINAL ULTRASONOGRAPHY
DURING FERTILITY EVALUATION
Wolman, Igal - Tel-Aviv, Israel
312.6 ULTRASOUND MONITORING AND HORMONAL PROFILES IN SERUM AND FOLLICULAR FLUID FOR
UNSTIMULATED IVF-CYCLES IN RELATION TO ÌEMPTY FOLLICLE SYNDROMEÎ.
Tang-Pedersen, Mikael - Odense, Denmark
312.7 A STUDY OF EPIGENETIC ALTERATIONS ASSOCIATED WITH PREMATURE OVARIAN FAILURE
Ghahremani, Manda - Vancouver, Canada
April 19-22, 2009 34

DETAILED PROGRAMME
WEDNESDAY, APRIL 22, 2009
08:30-10:15 Plenary III Room: Salle 2
Co-Chairs: Feki, Anis - Geneva, Switzerland

Jaconi, Marisa - Geneva, Switzerland
400.1 DERIVATION AND CHARACTERIZATION OF HUMAN EMBRYONIC STEM CELL LINES
Hovatta, Outi - Stockholm, Sweden
400.2 REPROGRAMMING SOMATIC CELLS: FUTURE PERSPECTIVES
Campbell, Keith - Loughborough, UK
400.3 TOOLS FOR STEM CELL RESEARCH AND CLINICAL APPLICATIONS: THE HUMAN EMBRYONIC STEM CELL
REGISTRY (HESCREG) AND THE ISSCR GUIDELINES FOR THE TRANSLATION OF STEM CELL RESEARCH
Veiga, Anna - Barcelona, Spain
10:45-12:30 Concurrent Symposium 13 - Controlled Ovarian Stimulation Room: Salle 3
Co-Chairs: Frydman, René - Clamart, France

Sallam, Hassan - Alexandria, Egypt
401.1 OUTCOME OF ASSISTED REPRODUCTION AFTER OVARIAN HYPERSTIMULATION AND AFTER
CRYOPRESERVED EMBRYO TRANSFER
de Geyter, Christian - Basel, Switzerland
401.2 OHSS PREVENTION AND TREATMENT
Sallam, Hassan - Alexandria, Egypt
401.3 MINIMAL STIMULATION IN IVF
Kato, Osamu - Tokyo, Japan
401.4 MATRIX METALLOPROTEINASE ACTIVITY DURING THE MENSTRUAL CYCLE
Abdallah, Michel Abou - Beirut, Lebanon
10:45-12:30 Concurrent Symposium 14 - Human ES and Reproductive Medicine Room: Salle 2
Co-Chairs: Jaconi, Marisa - Geneva, Switzerland

Feki, Anis - Geneva, Switzerland
Erenus, Mithat - Turkey
402.1 PROMISES AND CHALLENGES TOWARDS THE CLINICAL USE OF PLURIPOTENT STEM CELLS
Feki, Anis - Geneva, Switzerland
402.2 EMBRYONIC STEM CELLS DERIVED FROM PGD-AFFECTED EMBRYOS FOR MODELING HUMAN
GENETIC DISORDERS
Ben-Yosef, Dalit - Tel Aviv, Israel
402.3 THE STEM CELL NICHE IN THE TESTIS: POTENTIAL FOR REGENERATIVE AND REPRODUCTIVE MEDICINE
Vassena, Rita - Barcelona, Spain
35 Geneva
DETAILED PROGRAMME
10:45-12:30 Concurrent Symposium 15 - Surgery for Reproductive Medicine Room: Salle 4
Co-Chairs: Dubuisson, Jean-Bernard - Geneva, Switzerland

Nardo, Luciano - Manchester, UK
403.1 LAPAROSCOPIC TREATMENT OF UTERUS BICORNIS
van Herendael, Bruno J. - Antwerp, Belgium
403.2 DEEP PELVIC ENDOMETRIOSIS AND FERTILITY
Dubuisson, Jean-Bernard - Geneva, Switzerland
403.3 ROLE OF HYSTEROSCOPY IN INFERTILITY AND IVF
Mencaglia, Luca - Florence, Italy
403.4 REPRODUCTIVE SURGERY AND IVF
Gomel, Victor - Vancouver, Canada
14:00-15:45 Concurrent Symposium 16 - Egg Donation Room: Salle 3
Co-Chairs: Murdoch, Alison - Newcastle upon Tyne, UK

Boivin, Jacky - Cardiff, UK
404.1 SUCCESSFUL EGG DONATION PROGRAM
Murdoch, Alison - Newcastle upon Tyne, UK
404.2 OOCYTE SHARING AS TREATMENT OF INFERTILITY
Ahuja, Kamal - London, UK
404.3 OPTIMIZING TREATMENT WITH OOCYTE DONATION
Chillik, Claudio - Buenos Aires, Argentina
404.4 LUTHEAL PHASE SUPPORT, AN EVIDENCE-BASED APPROACH
Aboulghar, Mohamed - Cairo, Egypt
405.1 MECHANISM OF OOCYTE MATURATION

Jones, Keith T. - Newcastle, Australia
405.2 SELECTION OF PATIENTS AND OOCYTE COLLECTION FOR IVM
Holzer, Hananel - Montreal, Canada
405.3 PGD AND EMBRYO STATUS: ETHICAL AND MORAL ASPECTS
Dal Canto, Mariabeatrice - Monza, Italy
405.4 AMH/MIS AS A MARKER OF OVARIAN RESERVE AND PREDICTOR OF OVARIAN RESPONSE
Ledger, William - Sheffield, UK
April 19-22, 2009 36

DETAILED PROGRAMME
14:00-15:45 Concurrent Symposium 18 - Male Reproduction and ART Room: Salle 4
Chair: Palermo, Gianpiero - New York, USA

406.1 NEW GENETIC APPROACHES FOR THE SUCCESS OF ART
Benkhalifa, Moncef – La Verrière, France
406.2 IMPACT OF SPERM PREINCUBATION AND ACROSOME REACTION FOR ICSI OUTCOME
Mansour, Ragaa - Cairo, Egypt
406.3 HOW TO STUDY AND SELECT THE BEST SPERM FOR ICSI
Zorn, Branko - Ljubljana, Slovenia
406.4 MALE GAMETE EMPOWERMENT
14:00-15:45 Oral Communications 5 Room : Salle 18
Co-Chairs: Smith, Gary - Ann Arbor, USA

Demirol, Aygül - Ankara, Turkey
407.1 STUDY ON EFFECTS OF MALE REPRODUCTIVE TRACT INFECTIONS WITH AEROBIC BACTERIA
ON SPERM QUALITY
Kaluarachchi, Athula - Colombo, Sri Lanka
407.2 SEMINAL MORPHOLOGY AND THE OUTCOME OF ASSISTED REPRODUCTION TECHNIQUES
Kaluarachchi, Athula - Colombo, Sri Lanka
407.3 UTERINE SEPTUM DECREASES THE IMPLANTATION RATE IN IVF / ICSI CYCLES
Tomazevic, Tomaz - Ljubljana, Slovenia
407.4 A NOVEL PROCEDURE FOR SEVERE CASES OF ADENOMYOSIS – SURGICAL TREATMENT BY THE
TRIPLE-FLAP METHOD FOR RECONSTRUCTION OF THE UTERINE WALL
Osada, Hisao - Tokyo, Japan
407.5 PREVALENCE OF MARKERS FOR THROMBOPHILIA AND IMMUNOLOGICAL DISORDERS IN PATIENTS
HAVING IN VITRO FERTILISATION AND EMBRYO TRANSFER
Mettler, Liselotte - Kiel, Germany
407.6 ASSESSMENT THE OVARIAN RESERVE FOLLOWING CHEMOTHERAPY AMONG A COHORT OF CANCER
PATIENTS WHO UNDERWENT UNILATERAL OOPHORECTOMY VERSUS PARTIAL OOPHORECTOMY FOR
OVARIAN TISSUE CRYOPRESERVATION
Massarwa, Abir - Tel-Aviv, Israel
407.7 VASCULAR COMPLICATIONS OF OVARIAN HYPERSTIMULATION (OHS)
Dreyfus, Jean-Michel - Lyon, France
37 Geneva
DETAILED PROGRAMME
16:15-18:00 Concurrent Symposium 19 - AMH and Fertility Room: Salle 3
Co-Chairs: Remohí Gimenez, José - Valencia, Spain

Bischof, Paul - Geneva, Switzerland
Kadayifci, Oktay - Turkey
408.1 ANEUPLOIDIES IN OVARIAN STIMULATION
Remohí Gimenez, José - Valencia, Spain
408.2 BIOLOGY AND BIOCHEMISTRY OF AMH/MIS
Josso, Nathalie - Clamart, France
408.3 AMH/MIS AS A MARKER FOR THE PRESENCE OF TESTICULAR TISSUE AND AS A PARAMETER IN THE
WORK-UP OF MALE INFERTILITY
Fenichel, Patrick - Nice, France
408.4 AMH IN PCO PATIENTS
Fanchin, Renato - Clamart, France
16:15-18:00 Concurrent Symposium 20 - Psychological and Social Aspect Room: Salle 4

of ART Treatment
Co-Chairs: van der Poel, Sheryl - Geneva, Switzerland

409.1 LOW COST ASSISTED REPRODUCTION IN LOW-RESOURCE SETTINGS
van der Poel, Sheryl - Geneva, Switzerland
409.2 PSYCHOLOGICAL AND SOCIAL ASPECT OF ART TREATMENT
Verhaak, Chris – The Netherlands
409.3 SECURING GOVERNMENT FUNDING FOR IVF TREATMENT: THE CANADIAN EXPERIENCE
Hanck, Beverly - Montreal, Canada
409.4 PSYCHOLOGICAL COMPLEXITY SURROUNDING ENDING IVF TREATMENT
Takefman, Janet - Montreal, Canada
409.5 IMPACT OF STRESS ON IVF OUTCOME
April 19-22, 2009 38

DETAILED PROGRAMME
Co-Chairs: Jaconi, Marisa - Geneva, Switzerland

Aboulghar, Mohamed - Cairo, Egypt
410.1 EFFECTS OF L-GLUTAMINE AND STRAW SIZE, FREEZING RATE AND THAWING RATE UPON POST-THAW
QUALITY OF HUMAN SPERMATOZOA
Asherkaci, Hussian - Misurata, Libya
410.2 IN VITRO FERTILIZATION IN NATURAL CYCLE FOR CONCURRENT TRANSFER OF FRESH AND
FROZEN/THAWED EMBRYOS
Smirnova, Anna - Moscow, Russia
410.3 N-ACETYL-CYSTEIN IMPROVES RESULTS OF LONG-TERM CULTURE OF FROZEN / THAWED HUMAN
OVARIAN TISSUE
Fabbri, Raffaella - Bologna, Italy
410.4 VITRIFICATION OF DAY 3 EMBRYOS IMPROVES THE POST THAW PREGNANCY RATES
Pai, Hrishikesh - Mumbai, India
410.5 HIGH SURVIVAL AND PREGNANCY RATE AFTER SINGLE EMBRYO TRANSFER USING CRYOTOP
BLASTOCYST VITRIFICATION METHOD
Okubo, Tsuyoshi - Shinbashi Minato-ku, Japan
410.6 COMPARISON OF VITRIFICATION VERSUS SLOW COOLING FOR BLASTOCYST CRYOPRESERVATION
IN AN IVF PROGRAM
Dorfmann, Andrew - Fairfax, USA
410.7 FROZEN EMBRYO TRANSFER: THE EFFECT OF NUMBER TRANSFERD, STAGE TRANSFERD
AND SYNCHRONISATION ON THE CLINICAL OUTCOME
Salman, Tarique - Rochdale, UK
39 Geneva
NOTES
April 19-22, 2009 40

ORAL ABSTRACTS – MONDAY
Main References:
200.2 – NATURAL CYCLE IVF/M Steptoe PC, Edwards RG. Successful birth after IVF. Lancet 1978; 312: 366.
Jin-Ho Lim, Maria Infertility Hospital, Seoul, Korea. Lopata A, Brown JB, Leeton JF, et al. In vitro fertilization of preovulatory
The first live birth following in vitro fertilization (IVF) was produced from a oocytes and embryo transfer in infertile patients treated with clomiphene
natural cycle IVF (1). However, this was slowly replaced by ovary stimulated and human chorionic gonadotropin. Fertil Steril 1978; 30: 27-35.
IVF, because it was believed that the number of oocytes retrieved relates to Johnston I, Lopata A, Speirs A, et al. In vitro fertilization: the challenge of
the embryos available for transfer and this directly affects the probability of the eighties. Fertil Steril 1981; 36: 699-706.
successful pregnancy (2-4). At the beginning, the relatively inexpensive Jones HW Jr, Jones GS, Andrews MC, et al. The program for in vitro
clomiphene citrate was used to stimulate ovaries for producing multiple fertilization at Norfolk. Fertil Steril 1982; 38: 14-21.
follicles, but currently the ovarian stimulation protocols use the much more
expensive gonadotropin-releasing hormone (GnRH) agonist or antagonist in Beerendonk CCM, van Dop PA, Braat DDM, Merkus JMWM. Ovarian
combination with gonadotropin to generate multi-follicles in the ovaries. hyperstimulation syndrome: facts and fallacies. Obst Gynecol Surv 1998; 53:
439-449.
Some women are extremely sensitive to stimulation with exogenous
gonadotropin and are at an increased risk of developing ovarian Brinton LA, Moghissi KS, Scoccia B, Westhoff CL, Lamb EJ. Ovulation
hyperstimulation syndrome (OHSS), sometimes, a life-threatening condition induction and cancer risk. Fertil Steril 2005; 83: 261-271.
(5). In addition, there is an anxiety that the long-term side effects of Nagund G, Waterstone J, Bland JM, et al. Cumulative conception and live
repeated ovarian stimulation may increase the risk of ovarian, endometrial birth rates in natural (unstimulated) IVF cycles. Hum Reprod 2001; 16: 258-
and breast cancers (6). Although these problems are not encountered in 262.
natural cycle IVF treatment, a number of other problems arise, including an
Lukassen HGM, Kremer JA, Lindeman EJM, et al. A pilot study of the
increased risk of no oocyte retrieved during collection and no embryo
efficiency of intracytoplasmic sperm injection in a natural cycle. Fertil Steril
available for transfer. Nevertheless, there is resurgence of interest in natural
2003; 79: 231-232.
cycle IVF treatment in recent years, because the efficiency of IVF technology
has been improved markedly (7,8). Chian RC, Buckett WM, Tulandi T, Tan SL. Prospective randomized study of
human chorionic gonadotrophin priming before immature oocyte retrieval
Immature oocyte retrieval followed by in vitro maturation (IVM) of these
from unstimulated women with polycystic ovarian syndrome. Hum Reprod
oocytes is an attractive infertility treatment for women with infertility. In
2000; 15: 165-170.
comparison with ovary stimulated IVF treatment, the major advantages of
IVM treatment include avoidance of the risk of OHSS, reduced cost, and Buckett WM, Bouzayen R, Watkin KL, et al. Ovarian stromal echogenicity in
simplified treatment. Immature oocyte retrieval followed by IVM of these women with normal and polycystic ovaries. Hum Reprod 1999; 14: 618-621.
oocytes was initially shown to be a successful treatment for infertile women Chian RC, Buckett WM, Abdul Jalil AK, et al. Natural-cycle in vitro
with polycystic ovary syndrome (PCOS), because there are numerous antral fertilization combined with in vitro maturation of immature oocytes is a
follicles within the ovaries in this group of patients (9). Immature oocyte potential approach in infertility treatment. Fertil Steril 2004; 82: 1675-1678.
retrieval followed by IVM might be useful in up to approximately 30% of
Lim JH, Park SY, Yoon SH, et al. Combination of natural cycle IVF with IVM
women undergoing IVF treatment who have large numbers of antral follicles
as infertility treatment. In Chapter 27: In-vitro Maturation of Human Oocytes,
including patients with PCOS (10). Additionally because of the low cost of
basic science to clinical application edited by SL Tan, RC Chian, WM
therapy, it is important to identify candidates for IVM technology for women
Buckett. Informa Healthcare Press, London, UK. 2007; Pp 353-360.
with various causes of infertility. This strategy has been explored using
natural IVF combined with IVM for women without PCOS (11-13). Lim JH, Yang SH, Chian RC. New alternative to infertility treatment for
Therefore, it is important to evaluate the efficiency of natural cycle IVF women without ovarian stimulation. Reprod BioMed Online 2007; 14: 547-
combined with IVM as a clinical treatment for infertile women. 549.
As the protocol, the first line is to select the patients for natural cycle IVF/M
treatment for all patients who came to our infertility clinic, if a total of more 200.3 – ONCOFERTILITY: THE PRESERVATION OF FERTILITY OPTIONS FOR
than 7 follicles existed in both sides of ovaries at baseline ultrasound scan YOUNG PEOPLE WITH CANCER
on day 3-5 of the menstrual cycle. The second ultrasound scan was
performed on day 7-9 to ensure the size of follicles and endometrium Teresa K. Woodruff, The Feinberg School of Medicine, Northwestern
thickness. Human chorionic gonadotropin (HCG; 10,000 IU) was given when University, Chicago, IL, USA.
the size of the leading follicles reached to 12-14 mm in diameter and the Cancer is now a disease with a variety of treatment options, which are
thickness of endometrium is >6 mm, and oocyte retrieval was performed 36 leading to longer and more productive lives by survivors. Globally, there are
hours later. The leading and the small follicles were aspirated by 10 million people diagnosed with cancer. 10% of these newly diagnosed
transvaginal ultrasound-guided needle connected with aspiration pump. men and women are under the age of 45 years old. Infertility can be a
After oocyte collection, the mature oocytes from the leading follicles were consequence of many of the more aggressive chemo- and radiation
subjected insemination by intracytoplasmic sperm injection (ICSI) therapies that prolong and save lives. The ability to easily preserve sperm
immediately, and the immature oocytes were transferred to IVM-Medium for prior to cancer treatment provides hope at the time of diagnosis and families
culture. The immature oocytes were assessed for its maturity 24 hrs after later in life for male survivors. A notable example is Tour de France winner
incubation. If any oocytes matured, they were inseminated by ICSI. The Lance Armstrong who has three children conceived using sperm frozen days
resulted embryos from each patient were pooled together for transfer on day before he underwent the massive chemo- and radiation therapy that saved
3-4 following oocyte retrieval. For the preparation of the endometrium, the his life. Unlike sperm, the female germ cell, the oocyte or egg must be
patients were given estradiol daily starting on the day of oocyte retrieval. retrieved surgically. Moreover, the vast majority of collected oocytes will be
Luteal support was provided progesterone daily starting on the day of ICSI. immature and cannot be used immediately by a woman who is ready to start
We obtained approximately 40% clinical pregnancy rate, and more than a family. The overall hypothesis of the program is that effective fertility-
50% of patient population can be treated with natural cycle IVF/M or IVM extending options can be provided to young women undergoing
alone, indicating that natural cycle IVF/M is an efficient alternative of life-preserving cancer treatment. The purpose of our work is to bring
infertility treatment. We propose the new concept, namely, natural cycle physicians, medical ethicists, social scientists and basic scientists together
IVF/M, which is an efficient infertility treatment for all indications, especially to develop new strategies for fertility preservation for female cancer
for women under 35 years of age. In conclusion, the results from our data survivors under the new discipline of oncofertility. And even as the lexicon is
demonstrate that natural cycle IVF/M together with IVM alone can be used being established, complex bioethical issues face both providers and
to treat more than a half of population of infertile patients with acceptable parents. At the basic science level, complex issues of ovarian function and
pregnancy rate. preservation must be addressed including the problem of follicle growth and
development in vitro. Our investigative group has pioneered the development
of a 3-dimensional system that supports follicle development, largely, we
believe, because the links between the egg and its surrounding cells are
41 Geneva
maintained. Using a tissue-engineered approach, we have developed an in Conclusions: In an unselected population of patients undergoing IVF, a
vitro follicle growth system that supports the maturation of the enclosed substantial share (66.7 %) demonstrates pathological assays for
oocyte, which can be fertilized and results in live, healthy and reproductively thrombophilia or immunological disorders without symptoms of a clinically
competent mice. The goal of our program and the broader Oncofertility active disease. Every fourth (25.2 %) patient displays combinations of these
Consortium is to explore and expand the reproductive options available to laboratory parameters. These patients show a strong tendency towards
young people facing a fertility-threatening but life-preserving cancer decreased pregnancy rates after IVF.
treatment.
This work is supported by NIH/NICHD Structure-Function Relationships in 205.1 – ETIOLOGICAL FACTORS AND CLINICAL COURSE IN PATIENTS WITH
Reproductive Science, U54 HD041857; and, the Oncofertility Consortium, PREMATURE OVARIAN FAILURE
UL1DE019587 and RL1HD05829 http://oncofertility.northwestern.edu/
Bunpei Ishizuka, Saint Marianna University School of Medicine, Kanagawa,
Japan.
202.1 – FERTILOSCOPY BEFORE ART: A COMPLEMENTARY TOOL In an analysis of 130 consecutive cases with spontaneous premature
A. Watrelot, J.M. Dreyfus. Centre de Recherche et d’Etude de la Stérilité ovarian failure (POF), we found X chromosomal abnormalities in 15% and
(CRES®), Lyon, France. FMR-1 premutation in 2.5% of the cases. About half of the cases showed at
We have described the technique of fertiloscopy in 1997. least one type of autoantibody tested and about half of the cases with
positive autoantibodies had clinical autoimmune diseases.
Since then , we have demonstrated its efficiency, and its accuracy compared
to laparoscopy in a randomized prospective manner (the FLY study- Hum; In the present study, we analyzed the clinical course of POF of these patients
Reprod.2003,18:834-839). with different etiological factors.
After performing more than 2000 procedures, it seems now obvious that
Fertiloscopy practised early in the infertile work-up represent a valuable tool 205.3 – REGULATION OF GRANULOSA CELL APOPTOSIS BY GnRH
to decide which kind of ART will be the most effective in infertile patients Peter C.K. Leung, University of British Columbia, Vancouver, BC, Canada.
with no obvious pathology.
In view of the wide applications of GnRH analogs in assisted human
If every thing is normal including the tubal mucosa, then Intra Uterine reproduction, it is important to understand how GnRH exerts its effects in
Insemination (IIU) is the preferred option. When there is no pelvic different tissues. The expression of GnRH-I, GnRH-II and their common
pathology, systematic salpingoscopy and microsalpingoscopy allow receptor (GnRHR) has been demonstrated by immunohistochemistry in the
showing abnormal tubal mucosa in 22% of cases even if the tubes seem human ovary. Our analysis of the 5’-flanking regions of the human GnRHR
patent and macroscopically normal. In these cases, In vitro fertilization has gene has indicated a differential usage of specific promoter regions in
to be proposed directly. various tissues. Mutational analysis demonstrates the importance of three
Also in selected cases, reconstructive surgery per laparoscopy or per closely spaced CCAAT/enhancer binding protein and GATA motifs, which
Fertiloscopy may be offered if the tubal mucosa is normal function cooperatively in regulating GnRHR gene transcription in human
Therefore, using this mini-invasive approach, the best option is chosen granulosa cells. This granulosa cell-specific GnRHR promoter region further
without delay which decreases the time for patient to conceive supports the importance of an autocrine GnRH-GnRHR system in the
control of ovarian follicle development in humans.
Recent studies in our laboratory have demonstrated the direct effect of
204.4 – HIGH PREVALENCE OF MARKERS FOR THROMBOPHILIA AND GnRH-II in regulating ovarian steroidogenesis. Like GnRH-I, treatment of
IMMUNOLOGICAL DISORDERS IN PATIENTS HAVING IN VITRO human granulosa cells with GnRH-II results in down-regulation of hCG-
FERTILIZATION stimulated progesterone production. This inhibitory effect of GnRH-II can
L. Mettler, A. Salmassi, A. Schmutzler. Department of Obstetrics and be abolished by the GnRH-I antagonist Antide, suggesting a common
Gynaecology – Campus Kiel, University Clinics Schleswig-Holstein, Kiel, receptor. Similar to GnRH-I, GnRH-II does not interfere with the hCG-
Germany. stimulated cAMP production in the granulosa cells. Rather, these hormones
down-regulate the mRNA levels of FSH and LH receptors. Treatment with
Synopsis: In spite of the current disagreement among reproductive
GnRH-I produces a biphasic response in its own mRNA level such that high
immunologists, the study showed a complex pattern of coagulation and
concentrations decrease the GnRH-I mRNA level, whereas low
immunologic disorders impairing the outcome of IVF (66.7% of patients had
concentrations increase GnRH-I gene expression. In contrast, treatment
pathological assays).
with GnRH-II results in a homologous down-regulation of its mRNA level at
Objective: As birth rates after IVF remain low at 26 %, the hypothesis of the all concentrations. The expression of GnRHR is differently regulated by
high incidence of subclinical autoimmune and hematologic abnormalities in GnRH-I and GnRH-II. Also, the expressions of GnRH-I and GnRH-II are
IVF populations [1] is investigated. regulated differentially by FSH and hCG, such that the gonadotropins
Methods: In a first limited retrospective study we analysed 123 unselected increase the mRNA level of GnRH-II but decrease that of GnRH-I in hGCs.
patients undergoing IVF. Blood samples were determined using a selected Both GnRH-I and GnRHR mRNA levels have been shown to be down-
pattern of hereditary (protein-S, -C-deficiency, factor V Leiden), acquired regulated by estradiol . By contrast, estradiol increased GnRH-II mRNA
thrombophilia (antiphospholipid antibodies) and immunological parameters levels in granulosa cells.
(ANA, complement factor 4). We tested for only one antiphospholipid We have further observed that GnRH-I and -II induce apoptosis in human
antibody although we are aware that other APAs may, in fact, be more granulosa cells through GnRH-I receptors, which mediate the proteolytic
prevalent that ACA-Abs. caspase cascade involving caspase-8 (the initiator) and caspase-3 and 7
Results: No patient displayed clinical evidence for an active immunological (the effectors). GnRH-I and II induced TUNEL-positive apoptotic cells and
or hypercoagulative disease. Prevalence of protein S deficiency (11.2 %), increased cleavage activities of caspase-8, 3 and 7 by 48 hours and peaked
positive antiphospholipid antibodies (32.5 %) and ANA (30 %) were at 72 hours. Antide effectively blocked these TUNEL-positive changes and
significantly higher than in the normal population (< 1 %;p < 0.005; ~2 %;p expression levels of caspase-3 induced by GnRH-I or II. Activation of
< 0.005; ~5 %;p < 0.005). Overall 82/123 (66.7%) patients were positive for caspase-8, 3 and 7 was inhibited by the corresponding caspase inhibitor.
any pathological marker, 31/123 (25.2%) patients had a combination of Caspase-8 inhibitor also abolished cleavages of caspase-3 and 7 induced by
parameters of hereditary and/or acquired thrombophilia and/or GnRH-I and II. Interestingly, FSH protects human granulosa cells from
autoantibodies. Women with multiple abnormal tests were found to have a apoptosis induced by GnRH I or II. This raises potentially important roles of
strong tendency towards a lower pregnancy rate after IVF, although the GnRH-I and GnRH-II, together with FSH, in regulating ovarian follicle
difference did not reach statistical significance (38.7 % vs. 51.2 % development and atresia.
respectively; p > 0.005). [Supported by the Canadian Institutes for Health Research]
April 19-22, 2009 42

The accuracy of the different zygote features to predict implantation was

206.1 – HIGH SURVIVAL AND DEVELOPMENTAL RATES OF VITRIFIED MOUSE determined by ROC analysis. The area under the ROC curve (AUC) was used to
ZYGOTES FOLLOWING POLAR BODY BIOPSY determine the global performance of each feature to predict implantation.
E. Macas, X. Min, R. Stiller, S.G. Merki-Feld, P. Pelczar, B. Imthurn. Division of Results: Zygote images coming from the two IVF centres were successfully
Reproductive Endocrinology, University Hospital Zurich, Zurich, Switzerland. analysed with the software, resulting in a series of precise measurements of 24
variables for each zygote. Following this analysis, the only feature presenting a
Introduction: Pre-implantation genetic diagnosis (PGD) on polar bodies is one of significant difference between implanted (n=99) and non-implanted (n= 251)
the most technically complex procedures within the field of assisted reproductive zygotes was nucleolar precursor bodies (NPB) distribution quantified by
technology. Numerous specific problems can arise during any phase of this measuring the mean distance between NPB and the line separating the two
procedure, resulting in either an incomplete genetic diagnosis or none at all. For pronuclei (13.37±3.4 microns versus 14.39±3.03 microns, p=0.001).
instance, the actual biopsy itself poses serious technical difficulties, and even a Subjectively scored NPB distribution was also significantly different between
small error made here may lead to the loss or damage of the polar bodies. implanted and non-implanted zygotes (2.08 ± 0.56 versus 1.89±0.59, p=0.008).
Subsequent factors such the extensive micromanipulation of the polar body According to ROC, the computer-assisted measured NPB distribution presented
required during hypotonic treatment and fixation, as well as errors made during an AUC of 0.61 (95% CI 0.56-0.66) that indicates a poor prognostic efficiency of
fluorescence in-situ hybridization, may also seriously affect the safety and this parameter. However, all the other parameters presented lower AUC values. At
outcome of the entire biopsy procedure. Because of these problems, the need for an optimal threshold value (≤ 11 microns), specificity was high (88%) but
an effective freezing programme is particularly important in order to avoid sensitivity was low (33%), indicating that implantation was correctly predicted for
repetition of polar body biopsy if the excess zygotes are found to be only 33% of implanted zygotes. Compared to computer-assisted measurement,
chromosomally normal. However, studies conducted on biopsied zygotes subjective scoring of NPB distribution resulted in a lower AUC (0.58, 95%CI 0.52-
following application of conventional slow-freezing protocols are limited, and the 0.63) and a significantly lower sensitivity (20 %, p=0.000) at a specificity of 87%.
few results obtained thus far have been rather disappointing.
Conclusions: The morphology of human pronuclear zygotes can be objectively
In the present study, therefore, before introducing it into routine practice of the characterised by using an advanced image analysis tool. Among all the features
Division of Reproductive Endocrinology, University of Zurich, cryopreservation by analysed, NPB distribution appears as the most useful marker to predict
vitrification was tested using a mouse zygote. Applying this model the effect of implantation. However, its predictive value remains limited, probably because
two different methods of polar body biopsy followed by vitrification on the other factors, not related to zygote morphology, play a critical role in embryo
survival and development to blastocyst stage was examined. development and implantation. When embryo selection cannot be performed for
Material and Methods: Prior to vitrification, a total of 119 and 124 mouse legal constraints, the choice of zygotes on the basis of the distribution of their
zygotes were subjected to polar body biopsy using either laser-assisted or partial NPB remains an alternative that contributes to increase the implantation rates
zona dissection (PZD) techniques, respectively. Vitrification was also applied to after fresh transfer.
122 zona-intact zygotes that served as a control group.
Results: Following vitrification, no differences in the rate of zygote survival 206.3 – MATURE OVARIAN FOLLICLES CONTAIN A SUBPOPULATION OF STEM
(95.8%, 91.9% and 94.2%) or in the rate of development to expanded blastocyst CELLS
stage (82.3%, 79.8% and 81.9%) were observed between the two groups of
biopsied zygotes, or between the biopsied zygotes and control zygotes. The mean K. Kossowska-Tomaszczuk1, H. Zhang1,2, A. Scherberich2, I. Martin2, Ch. De
total number of cells comprising the blastocysts of controls (77.1 ± 4.7) was Geyter1,2. 1Woman’s Hospital; 2Department BioMedicine ; University Hospital of
comparable to the mean cell number recorded in the laser (66.4 ± 4.7) and PZD Basel, Basel, Switzerland.
(69.7 ± 5.3) groups. Blastocysts developed from laser-treated zygotes hatched Introduction: Luteinzing granulosa cells (GCs) reside in mature Graafian follicles
much earlier than blastocysts developed from the control and PZD groups of and are considered to be terminally differentiated destined to undergo apoptosis
zygotes (P < 0.001). within several days. We now demonstrate that a subpopulation of cells collected
Conclusion: The data obtained in the present study demonstrate that, from Graafian follicles possess all characteristics of multipotent stem cells.
irrespective of the biopsy method used prior to vitrification, mouse zygotes Material and Methods: Luteinizing GCs, collected from infertile women treated
survive and develop to blastocysts upon warming in proportions similar to those with assisted reproduction, were identified and sorted with a fluorescence-
of non-biopsied zygotes. activated cell sorter (FACS) based upon the presence of the follicle-stimulating
hormone receptor (FSHR). These sorted cells were then cultured in a culture
medium either supplemented with the leukemia inhibing factor (LIF) or not.
206.2 – COMPUTER-ASSISTED ANALYSIS OF HUMAN PRONUCLEAR ZYGOTES
Results: Luteinizing GCs cultured in the absence of LIF invariably became
F. Urner1, A. Beuchat2, C.O.S. Sorzano2, P. Thevenaz2, M. Unser2, T. Ebner3, A. apoptotic after approximately 10 days. However, in the presence of LIF luteinizing
Chanson1, M. Germond1, A. Senn1. 1FABER Foundation and Centre of Medically GCs remained viable for up to three months and obtained many of the
Assisted Procreation, Lausanne; 2Biomedical Imaging Group, Swiss Federal characteristics of multipotent stem cells, as given by the expression of various
Institute of Technology of Lausanne; 3Women’s General Hospital, IVF-Unit. Linz, surface markers such as CD29, CD44, CD90, CD105, CD117 and CD166. During
Austria prolonged culture in the presence of LIF the sorted cells progressively lost their
Introduction: To decrease the number of multiple pregnancies without affecting ability to express both FSHR and aromatase. In contrast, luteinizing GCs
pregnancy rates after in vitro fertilisation, a low number of embryos with continued to express OCT4, a marker of multipotent stem cells, but not vasa,
presumably high chances of implantation is usually transferred. In this situation, nanog nor stellar, markers of the germ-stem cell line. The multipotency of these
the accurate prediction of implantation is crucial. The assessment of zygote cells was then demonstrated by their differentiation into the neuronal,
morphology is one of the available tools to predict implantation when embryo osteoblastic and chondrogenic lineages when incubated in various specific
selection is not permitted, as in Switzerland. The aims of the present study are to culture media. In addition, we were able to demonstrate that multipotent stem
provide a software tool able to objectively measure several morphological cells developed from luteinizing GCs in the presence of LIF grew into
features of the pronuclear zygote and to identify which of these features are chondrogenic and osseous tissue after transplantation into the back of immuno-
useful to predict embryo implantation. incompetent nude mice.
Materials and Methods: To objectively analyse pronuclear zygote digital images, Conclusion: For the first time we were able to demonstrate that mature Graafian
a computer-assisted method was developed by creating a plug-in of the image follicles contain a subpopulation of multipotent stem cells. This novel finding may
processing ImageJ (http://rsb.info.nih.gov/ij). Images of zygotes were captured at have important implications for current concepts on follicular recruitment, the
the pronuclear stage 16-20 h after insemination under Hoffman contrast by using development of ovarian endometriosis and the pathogenesis of ovarian cancer.
an Octax camera (MTG, Herborn, Germany). Images of 350 pronuclear zygotes,
corresponding to transferred embryos with known individual implantation
outcome, were provided by two different IVF centres (Lausanne and Linz). They
were then analysed by using the ImageJ plug-in and by subjective scoring as
described previously by Senn et al. (Human Reproduction 21 :234-239, 2006).
43 Geneva
strongest transcriptional upregulations was observed for glycodelin A (PP14)

206.4 – INTERMEDIATE AND PREMUTATION FMR1 ALLELES IN WOMEN WITH which is known to be a progestagen-dependent, immunosuppressive, secretory
OCCULT PRIMARY OVARIAN INSUFFICIENCY epithelial endometrial product with high serum levels towards the end of the
luteal phase. Later the same technique was used to compare gene expression
I. Streuli, T. Fraisse, V. Ibecheole, I. Moix, M. Morris, D. de Ziegler. Hôpitaux profiles from endometrium after controlled ovarian hyperstimulation with tissue
Universitaires de Genève et Centre Hospitalier Universitaire Vaudois, Geneva, obtained in natural cycles. Again, differential expression was found for >200
Switzerland. genes, and it was concluded that current ovarian stimulation for IVF treatment
Introduction: The FMR1 premutation is now an established cause of primary was far from optimal. Gene expression profiles in patients with recurrent
ovarian insufficiency (POI) and it is currently recommended to test women with miscarriage in comparison to ongoing, uneventful pregnancy are not well
overt ovarian insufficiency. These recommendations led infertility specialists in investigated to date. The aim of this study was to differentiate the transcriptional
Geneva to request FMR1 testing more frequently from early 2005. Forty-four regulation of genes in the endometrium of patients with recurrent implantation
women were referred by infertility specialists for FMR1 testing between January failure (IF) versus those who became pregnant after IVF treatment, and
2005 and September 2006. Most women referred had signs of occult ovarian particularly between miscarried (M) and ongoing clinical pregnancies (OP).
insufficiency (oPOI, defined as FSH levels >10 UI/L, AMH levels < 7 pmol/L and/or Moreover, the effect of embryo-derived factors on endometrial transcriptional
a resistance to ovarian stimulation in infertile women with either regular or activity was studied.
irregular cycles under the age of 40 years) but did not meet the criteria for overt Material and Methods: Nine women with known IVF outcome (IF,M, OP) and
POI. Whether women oPOI are at risk of carrying expansions in the FMR1 gene is undergoing hysteroscopy were enrolled. An endometrial tissue sample was
currently not clear. biopsied using a soft suction curette. Mean patient age was 33.7 ± 4.5 years
The primary aim of our study was to establish whether intermediate and without difference between the groups. All tissue samples were taken during the
premutation alleles are more frequent in the population of infertile women with midluteal phase (day 23.6 ± 1.8 of a 30.0 ± 1.6 day cycle). Explant suspension
oPOI than in a control population. cultures were set up in order to maintain an intact three-dimensional
environment. After culture in presence of embryo-conditioned IVF media, total
Material and Methods: We compared the incidence of upper normal,
RNA was extracted using a method which included on-column DNAse treatment.
intermediate and premutation FMR1 alleles in women with occult primary ovarian
Three micrograms of total RNA per culture condition were submitted to reverse
insufficiency (oPOI) and in controls. In the study group we included 27 women
transcription, target cDNA synthesis, biotin labelling, fragmentation, and
with oPOI and a normal karyotype referred by infertility specialists for FMR1
hybridisation using the Affymetrix Human Genome U133A 2.0 Chip which covers
testing in 2005-2006 because of unexplained poor response to COH or altered
18,400 transcripts and variants including 14,500 characterised genes. Using the
hormonal profiles. In the control group we included 32 women undergoing
automated ranking of differential expression and their relevance in reproductive
genetic testing for conditions unrelated to mental retardation or ovarian function.
function, genes were selected for specific investigation with quantitative, real-
Results: In the group of women with oPOI, 4/54 alleles were in the upper normal time polymerase chain reaction (PCR) using the Taqman assay-on-demand
range (35-40 repeats), 4/54 were in the intermediate zone (41-60 repeats) and protocol. Results were calculated as threshold cycle (Ct) differences between the
3/54 were premutated (61-199 repeats). In the control group, 1/64 allele was in IVF outcome groups and normalised to GAPDH and -actin.
the upper normal range, 1/64 in the intermediate range and none was
Results: Automatic screening of the raw hybridisation data identified the genes
premutated. We compared the prevalence of alleles with 41-199 repeats and
which were found to be differentially up- or downregulated by a factor of 2 or
alleles with 35-60 repeats between the groups. The differences were statistically
more between the different IVF outcome conditions. When comparing endometrial
significant with p=0.02 et p=0.015 respectively.
tissue from OP with M, 1875 up- and 1807 downregulated genes were returned.
The analysis was also conducted at the level of individuals. One woman carried Real-time PCR analysis confirmed upregulation for somatostatin, PLAP-2, mucin
an intermediate and a premutation allele, and another carried two upper normal 4, and CD163, and downregulation of glycodelin, IL-24, CD69, leukaemia
range alleles. Therefore, in the oPOI group, 7/27 women had alleles ranging from inhibitory factor, and prolactin receptor. When the different embryo conditioned
35-60 repeats and 6/27 alleles ranging from 61-199 repeats. In the control group media were compared, no significant differential regulation could be
1/32 woman had an upper normal range allele and 1/32 an intermediate allele. demonstrated.
There is a relative risk of 7 of having an allele with 41-199 repeats (intermediate
Conclusions: Microarray profiling may currently not be sensitive enough for
and premutation range) in women with oPOI compared to controls with p=0.037.
studying the effects of embryo-derived factors on the endometrium. But given the
At the individual level, the difference between groups for alleles between 35-60
observed differences between M and OP, we believe that it will become an
repeats is not significant with p = 0.065.
interesting tool for the identification of fertility and obstetrically relevant markers
Conclusion: The results of our study show an increased incidence of alleles in produced by the endometrium in natural and stimulated cycles.
the intermediate to premutation range in patients with oPOI compared to the
general population. For this reason, the increased screening in Geneva seems
justified. In light of our findings, further efforts should be put into studying the 207.1 – DETECTION OF OXIDATIVE DNA DAMAGE BY FLOW CYTOMETRY AND
incidence of FMR1 expansions in all forms of ovarian insufficiency including ITS ASSOCIATION WITH MALE INFERTILITY
otherwise unexplained infertility. We suggest that FMR1 testing has a role in the Chakroun Feki Nozha1, Zribi Nassira1, Eleuch Henda2, Gdoura Radouane3,
comprehension of the aetiology of ovarian insufficiency and in the prevention of Sellami Afifa1, Bahloul Ali4 Hammami Adnene3, Gargouri Jalel2, Rebai Tarek1,
transmission of Fragile X syndrome and should be considered before infertility Keskes Ammar Leila1. 1Laboratoire d’Histologie Embryologie, Faculté de
treatments are initiated. Médecine de Sfax; 2Centre de transfusion sanguine, Sfax; 3Laboratoire de
Microbiologie, Faculté de Médecine de Sfax; 4Unité de recherche infertilité
masculine UR07US0011; Tunisie.
206.5 – GENE EXPRESSION PROFILING OF CULTURED ENDOMETRIUM FROM
WOMEN UNDERGOING IN VITRO FERTILISATION (IVF) WITH In the last two decades, the expanding research interest on the impact of
DIFFERENT OUTCOME sperm DNA damage on the ability of sperm to fertilize and on the pregnancy
outcome, has led to the development of various techniques evaluating
N.A. Bersinger, M.D. Mueller, M.H. Birkhäuser, D.M. Wunder. Klinik und
genome integrity of spermatozoa. DNA oxidation has been considered as
Polikliniken für Gynäkologie und Geburtshilfe, Universitätsfrauenklinik Inselspital
one of the important factors which affect sperm quality and increase the risk
Bern, Bern, Switzerland.
of genetic defects. Our aim with this work was to evaluate oxidative damage
Introduction: Successful embryo implantation is only possible during a short in human spermatozoa of infertile Tunisian men using the 8-oxo-guanine
time window within the luteal phase. Estradiol and progesterone are crucial for measurement by flow cytometry and to correlate results sperm DNA
the acquisition of receptivity and the change in transcriptional activity of target oxidation with semen parameters. Our work was performed on 15 semen
genes. Genomic microarrays have been used since 2002 to study the specimen coming from 15 infertile men; semen aliquots obtained by
implantation window. Comparing mid- to early luteal phase transcriptional masturbation were analyzed according to WHO guidelines. Leukocyte
activity in the endometrium a considerable number of genes was found to be concentration was determined by peroxidase staining using a
differentially expressed. For many of these a role in the context of endometrial heamocytometric method. We distinguished arbitrary 2 groups according to
receptivity or even fertility had not been previously described, but one of the the value of leucocytospermia [< 250000/ml (G1) and ≥ 250000/ml (G2)].
April 19-22, 2009 44

We measured sperm DNA oxidation by flow cytometry using the Oxi-DNA Routine serology screening was repeated in order to assess the suitability of
assay and correlated it with semen parameters and leucocytospermia. We his samples for patient treatment. The “signal response” to the HIV test was
used linear regression to correlate DNA oxidation, leukocyte concentration positive. A repeat test excluded a false positive result.
and semen parameters. Methods: Immediate action was taken to isolate all samples that may have
Interestingly, sperm DNA oxidation was correlated to leukocyte been in contact with the contaminated material. The HFEA were immediately
concentration in semen (p=0,006, r=0,7) and was significantly higher in G2 informed and the donor offered immediate consultation and counseling.
(p=0,03). These results suggest that low rates of leukocytes in semen There were 3 areas of concern, direct recipient infection, indirect
induce DNA oxidation and that the 8-oxo-guanine flow cytometry test can be contamination of adjacent samples and risk of infection of laboratory staff.
considered a sensitive biomarker of oxidative stress in human spermatozoa. A risk assessment of other samples within the same storage vessel became
important.
207.2 – IMPRINTING IN THE HUMAN EMBRYO: EVIDENCE FOR A RECYCLING Results: All stored vials from this donor were tested for HIV viral load. 4 of
OF HOMOCYSTEINE IN THE OOCYTE the 7 stored semen samples did express viral copies of 1 HIV gene or more.
However, the viral load was only significant in one sample. This sample was
Moncef Benkhalifa, Debbie Montjean, Paul,Cohen-Bacrie, Yves Menezo. the first stored immediately after the initial negative HIV serology test and
UNILABS, Laboratoire d’Eylau, Paris, Ile de france, France. yet 9470 viral copies/ml with 3 HIV genes were found.
Introduction: Assisted reproductive technology, especially with poor quality Within that storage tank there were samples from 34 different donors, of
sperm has been recently in the centre of controversies concerning which 3 were stored within the same canister as the HIV infected donor.
eprgenetic risks. Epigenetic plays an important role in many cancers that Random selection of these samples were also tested HIV viral load, using 4
appear late in life and may have roots in epigenetic variation established in methods. No HIV was detected. However, all the samples were discarded.
early development. One of the very early epigenetic regulations in life is DNA
methylation strongly involved in imprinting, catalysed by DNA Discussion: Investigation was complicated by the difficulty of obtaining clear
methyltransferase and requiring S-Adenosyl-Methionine (SAM) as a methyl advice regarding the contamination risk to staff and the frozen samples in
donor. Imprinting which also regulates implantation. Homocysteine (Hcy), the same dewar. Various scientific bodies and individuals were questioned
released after méthylation, is a well known inhibitor of methylation but without any contribution of value. Literature searches confirmed that
cross contamination of samples through liquid nitrogen has never been
Material and Methods: We have used microarrays, Affymetrix HG U-133 documented and this case emphasizes that contamination does not seem to
plus 2.0 chips, in order to determine the expression of the pathways occur.
removing or recycling homocystein, in 6 pools of GV oocyte collected at the
time of ICSI Although this was essentially a fact finding exercise we have modified our
storage arrangements to reduce the impact of a recurrence. This includes
Results: We confirm our previous data indicating that SAM synthesis is more diligent analysis of any change in sample quality and more regular
active: There is a high level of expression for methionine assessment of risk behavior. The HFEA were supportive and helpful at all
adenosyltransferase (MAT) II, alpha and MAT II beta. The S-Adenosyl- times. Their system of ‘alerts’ is recommended. We feel though that this
homocysteine hydrolase which releases homocysteine from SAH is strongly does not supersede the established scientific practice of early recourse to
expressed: this leads to high level of formation of Hcy post methylation. publication and dialogue.
All the enzymes involved in recycling of Hcy are highly expressed: 5, 10
methylene tetrahydrofolate reductase (MTHFR) and methionine synthase
(MTR). We found also that methionine can be form using the betaine 207.4 – PRESENTATION OF INNOVATIVE PATENTED PRODUCTS FOR OVUM
homocysteine pathway (BHMT2). However we were unable to detect any PICK UP AND EMBRYO TRANSFER
signal for cystathionine beta synthase (CBS), which allows recycling Hans-Peter Steiner, Institute for In-Vitro-Fertilization and Endocrinology,
homocysteine towards cysteine, contrarily to what is currently observed in Graz, Upper Austria, Austria.
animals. The following innovative devices for IVF/ET are products of the author`s 20-
Conclusions: If we consider the biochemical pathways present and years experience in IVF.
considering that at least a part of the mRNAs detected are translated, the The current worldwide ovum pick up technique in in vitro maturation (IVM)
human oocytes is able to keep the Hcy level in check via re-methylation. This is using a single lumen needle without flushing the follicles. In order to flush
aspect is important as recent works have shown that controlled ovarian the needle system accordingly repeated withdrawal and reinsertion of the
hyperstimulation affects the homocysteine concentration in the follicular needle into ovaries (up to 15- 20 times) was necessary so far. This
fluid. This recycling may alleviate, the imprinting inhibitory effect of Hcy; yet procedure is painful, time consuming and there is a high risk for
this can be modulated by maternal age and culture conditions it has to be contamination of the cumulus-oocyte-complex with blood. STEINER Needle,
mentioned here that these two methionine recycling pathways are also on the other hand, is 18 Gauge having a double lumen (starting 7cm
present in the testis proximal of needle tip) allowing for flushing of one follicle at a time within a
ADA: Adenosine de-aminase, AK: adenosine kinase, CBS: cystathionine beta few seconds without removal of the needle and, thus, minimizing the
synthase, MAT: méthionine adénosyltransférase, MTAses: various methylases contamination with blood. In addition, no filters are needed.
MTR: Methionine synthase, MTHFR: Methylene tetra hydrofolate reductase, In times of mild IVF and IVM flushing follicles has a revival. STEINER Flush
SAHH: SAH hydrolase is the first mechanical flushing pump worldwide activated by the physician
´s foot via a cable pull and it enables to choose the amount, velocity and
207.3 – HIV SERO-CONVERSION OF A SPERM DONOR DURING THE DONATION pressure of flushing medium.
PERIOD ETFLEX Steiner was particularly invented for difficult embryo transfers as
Venessa Smith, Geetha Venkat, Eric Simons, Shailaja Nair, Kamal Ahuja. well as mock Transfers (it has a cm scale). The metallic outer catheter with a
London Womens Clinic, London, UK. spiral tip (up to 5.5cm) can be used as a disposable or reusable device. The
spiral tip is clearly seen at ultrasound.
Background: An incident of HIV sero-conversion of a donor during the
donation period is documented. We also report the subsequent efforts made The STEINER Pistol enables the embryologist and physician to aspirate and
by our clinic to ensure the safety of patients and staff and to rule out other inject a reproducible amount of fluid during Embryo Transfer. Per click 5 or
samples which might have been exposed to the contaminated material. 10µl can be aspirated or transferred.
The recruitment policies of the donor bank are based on recommendations Short videos sequences and computer animations will complete the
by BAS, BFS, NICE, BICA and the HFEA Code of Practice. The sperm donor presentation.
successfully passed initial serum and STD screening and joined the program
in August 2005. During the subsequent 5 months his donations declined in
quality, as a result few were stored. His inclusion was then terminated.
45 Geneva
and after semen processing by Pure Sperm gradients centrifugation and its
207.5 – COMBINING TWO ZWITTERIONIC BUFFERS IN IVF HANDLING MEDIA effect on fertilization and pregnancy rate.
SUPPORTS MOUSE BLASTOCYST DEVELOPMENT AND NORMAL Material and Methods: Semen sample of patients (n= 87) attending our
HUMAN OOCYTE FERTILIZATION FOLLOWING ICSI clinic were collected from men undergoing IVF (n=36) or ICSI (n=51)
Jason E. Swain1, Virginia Ord2, Darleen Taylor2, Veronica Sossamon2, therapy after minimum 3 days of sexual abstinence. A routine semen
Thomas B. Pool2. 1Center for Reproductive Medicine, University of Michigan, analysis was performed according to WHO guideline (1999). Semen
Ann Arbor, MI; 2Fertility Center of San Antonio, San Antonio, TX; USA. samples were prepared using two layers (90/45) of PureSperm gradient
centrifugation. Each aliquot of liquefied semen was layered on top of the
Introduction: Maintenance of stable pH is important for optimizing gamete gradient and centrifuged at 250xg for 20 min. The sperm pellet was used for
and embryo culture. This is especially true when handling the denuded oocytes insemination or injection. Semen vitality was evaluated using Eosin
oocyte, which lacks robust pH regulatory mechanisms. One method to test and the membrane integrity was evaluated by performing Hypo osmotic
stabilize pH entails using zwitterionic buffers in handling media used outside swelling test (HOS-test). Besides, the DNA integrity of individual
the laboratory incubator. Current handling media utilize single buffers, such spermatozoa was determined using chromomycine staining CMA3 (Sigma).
as MOPS or HEPES. However, use of a single buffer limits the ability to Whereas, DNA stand breaks was evaluated using TUNEL-Kit (Hoechst,
adjust optimal buffering capacity. For example, the pKa, or optimal company). ROS was measured using a commercial kit (Randox; company).
buffering of MOPS at 37°C is 7.02, which is below the traditional pH range
used to handle oocytes and embryos of ~7.3. Furthermore, potential Results: Sperm vitality, membrane, DNA integrity and DNA stand breaks in
toxicity concerns exist with elevated concentrations of buffers, such as fresh semen samples were (41.9±24.2%; 57.6±21.5%; 44.4±24.3%; and
HEPES, required in mono-buffered handling media. Therefore, traditional 51.4±22.9% respectively) and the corresponding values after semen
handling media utilizing a single buffer may not be ideal. processing were (53.9±21.2%;69.8±18.8%; 64.5±20.8%; and
70.5%±18.9% respectively). In both groups ROS concentration in the native
Materials and Methods: Media was prepared, varying the type and amount semen samples was 46.4±67.86 µmol/l. ROS in seminal plasma correlate
of zwitterionic buffer. Media containing HEPES, MOPS, or HEPES+MOPS negatively, but not significant, with vitality and membrane integrity before
were tested. All other parameters, including pH and osmolarity were and after semen processing. Whereas, a significant negative correlations
controlled. An acid/base challenge, using incremental addition of 0.1M HCL were observed between ROS and DNA integrity (r=-0.295; p=0.006) and
and NaOH, followed by pH measurements, was used to examine buffering DNA Stand breaks (r= -0.253; p=0.008) not only before, but also after
capacity of various media. Additionally, blastocyst development and cell semen processing (r=-0.283; p= 0.008 and r=-0.213; p=0.047). Besides; a
number of frozen/thawed 1-cell mouse embryos grown in respective media negative correlation was shown between ROS concentration in seminal
at 37°C in room atmosphere was used to validate efficacy. Finally, plasma and fertilization and pregnancy rates (r=-0.095; p=0.382 and r=-
fertilization and embryo development following microinjection of human 0.125; p=0.267 respectively).
oocytes in our standard HEPES-buffered medium was compared to a
HEPES/MOPS-buffered medium. Participants included all patients (n=12) Conclusion: ROS concentration in seminal plasma affects spermatozoa’s
undergoing ICSI with at least 6 mature eggs. Half of the oocytes were quality but can not affect the fertilization and pregnancy rate in IVF/ICSI
injected in a 10mM HEPES-buffered Tyrodes medium. The remaining mature programme.
oocytes were injected in media containing a 1:1 ratio of MOPS/HEPES (5mM
of each). Treatment groups were alternated in respect to their order of 208.5 – CURRENT CRITICISM ON IVM TREATMENT
injection to avoid bias. All other conditions besides holding media used for
microinjection remained consistent between treatments. Data were analyzed Milton Leong, Hong Kong, China.
using Chi-Square. IVM is a laboratory technique initially used to rescue germinal vesicle
Results: We confirmed the ability of a double-buffered handling media, oocytes and make them suitable for fertilization. In 1968, Edwards was the
containing both HEPES and MOPS, to allow adjustment of pKa value and first to demonstrate IVM in mouse oocytes; and then applied to human
optimal buffering capacity compared to media containing HEPES or MOPS oocytes, and from research to treatment form in the ensuing decades.
alone. This allowed for the subsequent reduction of individual buffer However, it wasn’t until the end of the 20th Century that consistent
concentrations, while maintaining overall buffering capacity. This double- pregnancy rates were obtained, and probably not until the last 5 years that it
buffered media also yielded similar rates of mouse blastocyst development became an established clinical modality, largely through the efforts from
and total cell numbers compared to media containing only a single buffer. Scandinavia, Korean, Asian groups, and especially the works from Montreal
Though no significant differences exist, injection of human oocytes in Canada.
double-buffered media with HEPES/MOPS (n=69) tended to have greater One must not forget that IVM is not a stand-alone fertility treatment, but
normal fertilization (71.0% vs. 62.7%) and lower abnormal fertilization merely a technique to prepare the oocytes for fertilization, and that IVF-ET is
(4.3% vs. 11.9%) than oocytes injected in HEPES alone (n=67). No still involved. Strictly speaking, it should be termed IVM/IVF.
differences were apparent in rates of multi-nucleation on day 3 between
At the present day, a good, experienced center should be able to achieve and
MOPS/HEPES and HEPES alone (12.2% vs. 16.7%). Continued experiments
IVM rate of 70-80%, with similar fertilization rates. Pregnancy rates should
to increase patient number may allow data to reach significance.
reach 30-40% per cycle, if 3 embryos are replaced. There have been over
Conclusion: Use of a double-buffered handling medium for ICSI is at least as 2000 babies born in the world. These results make IVM an alternate
effective as a traditional HEPES-only buffered medium. This approach of technique to conventional ovulation stimulation and IVF, especially in the
combining multiple organic buffers to regulate media pH offers the potential patients suffering from PCOS as well as the hyper-responders. However,
advantages of reducing individual buffer concentration and resulting toxicity controversy exists as to whether IVM should be accepted as a valid
concerns, as well as the ability to fine-tune and optimize maximal buffering treatment for infertile couples, and whether it can be a truly universally
capacity at a specific pKa value at a specific temperature. applicable technique.
Criticism on IVM treatment will be discussed in the following headings:
207.6 – CORRELATION BETWEEN REACTIVE OXYGEN SPECIES (ROS) IN A. Technical:
SEMINAL PLASMA AND SPERM VITALITY, MEMBRANE, DNA i. ?Secret recipe?
INTEGRITY, DNA STRAND BREAKS OF INFERTILE MEN BEFORE AND ii. FSH priming
AFTER SEMEN PROCESSING BY PURESPERM AND IVF/ICSI OUTCOME iii. HCG loading
M.E. Hammadeh, C. Fischer-Hammadeh, A.S. Amer, M.F. Hamad, M. Deryal. iv. Culture systems
Department of Obstetrics and Gynecology, University of Saarland, v. Identifying oocytes
Homburg/Saar, Germany. B. Clinical:
Introduction: the purpose of this study was to determine the effect of ROS in i. choice of patients
seminal plasma of infertile men undergoing IVF/ICSI therapy on the sperm ii. FSH priming
vitality, membrane integrity, DNA integrity and DNA strand breaks before
April 19-22, 2009 46

iii. Significance of endometrial thickness

iv. ISCI – always? 211.2 – THE ROLE OF IMMUNOHYSTOCHEMISTRY IN COMPLEX
v. Egg collection techniques MORPHOLOGICAL DIAGNOSIS OF WOMEN STERILITY
C. Outcome: Irina N. Kostyuchek1, Sergey V. Nikitin2, M. Kleshchov1, Kiryl I.
i. inconsistent pregnancy rates Prashchayeu3, Viktoria O. Polyakova1 , N. Balashova1, Igor M. Kvetnoy 1. 1Ott
ii. Miscarriage rates Institute of Obstetrics and Gynecology Russian Academy of Medical
iii. Baby Registry and follow up birth defects and other effects Science, Saint-Petersburg; 2 Clinica «Andromeda», Saint-Petersburg;
iv. Epigenetic changes 3
Belgorod State University, Belgorod; Russia.
Conclusion: IVM is a valid technique giving more flexibility to clinical Introduction: Inflammation, endocrine disorders and resistance of
treatment of infertility, and also for those who desires fertility preservation. endometrial cells lossed hormonal receptors are the most common reasons
There are problems to be solved as long-term results are yet to be realized, of sterility. The aims of this study were to characterize estrogen and
and its benefit should be weighed against probably undesirable effects and progesterone receptor status , identify immune cells in endometrium and
to be used with prudence as with any other clinical modality compare this data with histological features of uterine mucosa in sterility
women.
209.3 – ISO STANDARDS FOR CLINICAL WORKUP - EXPERIENCES WITH DIN Materials and Methods: 16 endometrial biopsies from 21-41 years old
EN ISO 9001:2000 sterility women (middle age 30 yo) was investigated on 22-24 day of
Bruno Imthurn, Division of Reproductive Endocrinology, Department of menstrual cycle. After histological study immunohistochemical identification
Obstetrics and Gynaecology, University Hospital, Zurich, Switzerland. of ER, PR, CD4, CD8, CD16, CD57, CD20 and HLA-DR was carry out.
Quantitative analysis of immunohistochemical expression was done by
The introduction of a quality management (QM) system into an institution using computer image analisator «Video-Test, Morphology-5».
usually goes along with some scepticism on the part of staff. However,
although perhaps successful regarding pregnancy rates as a matter of fact Results: Chronical endometritis, which characterized by limphocytic
many centres suffer under a chaotic organisation, unstructured infiltration and stromal fibrosis of endometrium was observed in 6 cases.
administration and little defined training processes for their staff. CD16 and CD57 were positive in NK-cells, CD20 – in B lymphocytes, CD 4
and CD8 – in T lymphocytes. In 2 cases autoimmune inflammation was
Among the different QM systems DIN EN ISO 9001:2000 especially fits the probable (high level of HLA-DR positive cells and negative microbiological
needs of an ART clinic, as it is internationally acknowledged, successfully investigation of endometrium). In 6 biopses luteal deficiency were
applied by many European IVF centres and widely known by the public. observed.Morphologically it’s appears by disoders in endometrial secretory
The following main issues are covered by DIN EN ISO 9001:2000: 1. General transformation, decrease of PR expression and comparative increase of ER
procedures; 2. Patient satisfaction; 3. Clinical aspects; 4. Laboratory expression in endometrial glands and stroma. In one patient combined
methods; 5. Staff training and development. disease took place: chronical endometritis and luteal deficiency.
Inflammation and immature regression of corpus lutea was in one case.
Based on our own experiences the presentation intends to give other
Typical endometrial hyperplasia with increase expression of ER and low level
colleagues hints and tricks, how to introduce a QM system into an ART
of PR was in one biopsy.
centre.
Conclusion: Immunohistochemical methods accompaning classical
As a consequence DIN EN ISO 9001:2000 resulted in our unit (1. positive) in
histologocal examination elaborate causes of sterility and useful for choice
a more structured organisation, less improvisation and higher pregnancy
of treatment.
rates, but also (2. negative) in more paperwork and higher costs.
211.3 – WHAT WE OFFER TO WOMEN BEFORE UNDERGOING SURGICAL

211.1 – EFFECT OF SEASON ON FERTILIZATION AND EMBRYO QUALITY RATES
TREATMENT FOR MENORRHAGIA
IN ART
Kinza Younas, A. Khan, S. Younas, R. Settatree. Obstetrics & Gynaecology
M.A. Danfour, W.M Elmahaishi, H.M Asherkaci, O.A Elsraite, M.S
Solihull Hospital Heartofengland NHS Trust; Birmingham Heartlands
Elmahaishi. Faculty of Medicine – 7th October University and Misurata
Hospital, Birmingham, UK
Infertility Centre, Misurata, Libya.
Aims/Standards: To determine whether women with menorrhagia are
Several studies have investigated seasonal variations during IVF. As known
undergoing surgical treatment unnecessarily without trying all possible
causes for reducing the rate of success of in-vitro fertilization (IVF). Their
medical methods. To see the treatment outcomes of Mirena coil.
results are contradictory, especially concerning fertilization and pregnancy
rates. The aim of the present study was to re-evaluate these parameters Nice guideline for the management of heavy menstrual bleeding no.44
using a large number of IVF cycles. METHODS: A total of 2250 ICSI cycles Jan.2007. RCOG guidelines for the management of menorrhagia.
(first trial ICSI 1411) conducted in Misurata IVF Center, Misurata-Libya Background: Excessive menstrual bleeding, which interferes with women’s
between February 2002 and august 2006 were retrospectively analysed. To Physical, social, emotional and quality of life. In early 1990s 60% of women
avoid a bias in the evaluation of the fertilization rate, the second and third with menorrhagia underwent hysterectomy while this trend is declining now.
ICSI trial for some patients were considered for analysis. After adjusting for There are recommendations for the medical management of menorrhagia.
confounding by the age of the woman, type of infertility, indication for IVF
and year of aspiration, some seasonal variation was observed in the oocyte Methods and Materials: Random, prospective questionnaire survey at
maturation, oocyte quality, fertilization rate, embryo quality. RESULTS: There Solihull and Birmingham Heartlands Gynaecology unit, wards and the clinic.
were statistically significant seasonal differences in Libya (Misurata) that 52 completed questionnaires analysed.
influenced the outcome of IVF treatment. The highest fertilization and Results: Duration of menorrhagia with the mean 9 years, mode 3,4 years
quality-A embryo rates were observed during the spring and the lowest in and median 5 years. Mirena coil was offered in 41% of patients only and
the autumn. These changes correlated with the absolute number of light (7/21) 13% tried the treatment while 2 stopped using it because of side
hours and humidity and its increment over time, but not with the effects. Tranexamic acid was tried in 11(52%) while Mafenamic acid was
temperature. CONCLUSION(S): Seasonality seems to have a significant tried in 7 (13%). Endometrial ablation was tried in 11 cases (21%).
influence on the fertilization process and on the quality of the human Norethisterone was tried in 5 (10%). Mirena coil was offered by hospital
embryos that are obtained in vitro, possibly because of the light/dark doctor in 18 cases as compare to only 01 case by the GP.
variations over time and humidity. More studies required to confirm this
finding. Once it confirmed, these seasonal changes should be taken into Recommendations: •Primary care services should be encouraged to follow
account when evaluating infertility data and in everyday clinical practice. the guidelines by the NICE and RCOG. •Try all the available resources for the
management before the surgical options.
Conclusion:•General practice should be encouraged to offer the Mirena as a
principle treatment for menorrhagia. •More data is required to come to a
conclusion and re-audit the practice in due course of time.
47 Geneva
AZH. Meta analysis suggests that the probability of live-birth after fresh IVF
211.4 – GLYCODELIN ENHANCES FERTILIZATION DURING IVF is higher after blastocyst-stage embryo transfer compared to cleavage-stage
embryo but there are not recent studies specifically for women over 40.
Nadja Gneist, Gudrun Keck, Ina,Trinkaus, Birgit Leuchten, Katrin Hanseroth, Recent RCTs did not show an increase in live birth rate after PGS in relation
Evelyn Gouma, Wolfgang Distler; Klinik fur Frauenheilkunde und to advanced maternal age. In the first trial the implantation rate was greater
Geburtshilfe, Dresden, Saxony, Germany. in the PGS group (17.1% v 11.5%) but was statistically not significant.
Introduction: During the first day of IVF oocyte morphology and its Oocyte donation appears the most successful option for women with
developmental potential can not be evaluated as prognostic value, only the advanced reproductive age as the success rates do not alter with the
expansion of the cumulus oophorus complex (COC) gave a hint for the recipient’s age.
oocyte maturity. Previous investigations of our study group about Conclusions: Despite many women’s expectations, systematic review of all
Glycodelin (Gd) in cumulus cells derived from denudation of ICSI-cycles available evidence suggests that the Assisted Reproduction Technology
showed a higher Gd-level correlated to mature eggs. In this present study continues to have very low live-birth rates in women over 40. Analysis of
we investigated cumulus cells from IVF cycles for Gd and correlated the trials showed that assisted hatching may increase the chance of pregnancy
results first to oocyte and COC maturity, respectively, and second to the in women with poor history. The application PGS for selection of good
fertilization outcome, which was of special interest because of the assumed quality oocytes in elderly women have not increased the success rates. The
selective potential of the cumulus oophorus. As described in literature, management of the infertile couple over 40 needs to focus on the exclusion
cumulus cells are able to modify the uptaken exogenous Gd-isoforms GdA of other factors apart for age, such as male or tubal factor, as well as
and GdF to GdC; thereafter spermatozoa-zona pellucida binding becomes education on normal ageing and fertility and psychological support. An
stimulated. evidence-base approach will protect infertile couples form investigations or
Material and Methods: We investigated the morphology of 125 COC derived interventions that are not proven worthwhile. In terms of population strategy
from 21 patients undergoing IVF (6,0± 2,3 per patient) after follicular the key to the management of infertility in women over 40 is prevention.
aspiration. All patients gave signed informed consent. A compact COC was
valued as “immature”. During maturation the cumulus expanses, the
oocytes becomes visible. Expanded COCs have been valued as “mature”. In 211.6 – ROLE OF DIAGNOSTIC LAPAROSCOPY IN WOMEN WITH NORMAL
addition we defined a borderline stage, which corresponds to the beginning PELVIC IMAGING AND FAILED SUPER OVULATION AND INTRA
of cumulus expansion. Each of the COC’s have been grouped into one of UTERINE INSEMINATION
these stages. Kay Jayakrishnan, Aby Koshy, Raju, RKJK Hospital, Fertility Research and
16 till 18 hours after insemination we performed PN-scoring. The oocytes Gynaec Centre, Trivandrum, Kerala, India.
became denuded and were transferred into fresh culture media. The Introduction: Though the advent of laparoscopy had revolutionized the way
fertilization outcome has been assessed. The remaining cumulus cell- sperm we diagnose and treat women with infertility, there are areas of debate on
suspension has been prepared using cytospin centrifugation. The Gd- the role of diagnostic laparoscopy in infertility. Laparoscopy has long been
immunostaining and semiquantification followed. used in the evaluation of patients with infertility where other diagnostic
Results: Gd has been found in cumulus cells after IVF. The staining showed methods have failed to come up with a cause. It also has the advantage of
variations that were found being correlated to oocyte maturity of the being a see and treat modality. Though a considered a safe procedure,
corresponding oocyte. While the COC maturity increases, the Gd-level of the laparoscopy is associated with complications inherent to the surgical
cumulus cells rises (p<0,05). The PN-scoring showed a regular 2PN-pattern procedure and the anaesthesia administered and these risks should not be
being accompanied with an increased Gd-level of the COC (p<0,05). disregarded. The costs incurred and the risks involved with the procedure
needs to be balanced with the therapeutic benefit. Though improvements in
Conclusions: First, these results show a clear correlation of ooctye maturity imaging technologies have reduced the requirement for diagnostic
and cumulus Gd, and second the necessity of Gd in the COC. The isoform laparoscopy, it is still considered the gold standard for the evaluation of the
Gd C is described having a spermatozoa- zona pellucida binding enhancing pelvis. We therefore undertook a retrospective study over an 8 year period
capacity. It can be hypothesized, that only in a mature COC the interactions to evaluate the role of diagnostic laparoscopy and hysteroscopy in women
between the Gd-isoforms and the spermatozoa are advancing the with normal ultrasound findings who failed to conceive after 3 or more
fertilization process. cycles of super ovulation and intra uterine insemination (IUI).
Materials and Methods: A retrospective study of patients who underwent
211.5 – A SYSTEMATIC REVIEW OF MANAGEMENT OF INFERTILITY IN diagnostic laparoscopy between January 2001 and December 2008 was
WOMEN OVER THE AGE OF 40 performed. Those patients who had no detectable pathology based on
history, physical examination and 3 D ultrasound and had treatment for 3 or
Gerasimos Marinakis, Dimitrios Nikolaou. Imperial College of Science,
more cycles in the form of super ovulation and IUI were included in the
Technology and Medicine, Chelsea and Westminster Campus, Ovarian
study. Moderate and severe male factor infertility was excluded in the study.
Ageing and Fertility Clinic, London, UK.
All patients underwent diagnostic laparoscopy and hysteroscopy under
Introduction: The current systematic review looked at the latest data from general anaesthesia. Patient profile, intra operative findings and operative
national statistics on fertility trends in the 40s, as well as the most up-to- complications were noted. Statistical analysis was performed using SPSS
date results of IUI and IVF, and the effectiveness of additional interventions 16.
and adjuvant treatments in this age-group.
Results: 127 patients underwent diagnostic laparoscopy and hysteroscopy
Materials and Methods: Data on natural fertility in the forties were retrieved in the study period. The mean age of the patients was 29.45 ± 4.34 years.
from National Statistics bureaux from UK and USA, as well as Eurostat. The The mean duration of infertility was 5.09 ± 3.03 years and a mean of 4.96 ±
latest results of Assisted Reproduction Treatments were also retrieved from 2.4 cycles of super ovulation and IUI were performed prior to the surgical
the HFEA. Finally, the effectiveness of additional interventions such as AZH, procedure. There were no major intra or post operative complications.
blastocyst transfer and PGS, as well as various adjuvant treatments, was 12.6% (n=16) patients had normal findings on laparoscopy and
examined in published meta-analyses, as well as primary searches in hysteroscopy with bilateral patent tubes. 77.2 % had endometriosis (AFS -
MEDLINE and EMBASE minimal (37.8%, n=48), mild (33.1%, n=42) and moderate (6.3%, n=8)) and
Results: Natural fertility in the 40s has remained low despite all the changes 5.5% (n=7) had pelvic inflammatory disease (PID). One patient with PID had
in lifestyle and medical advances. There is a well documented steady tuberculosis diagnosed on histopathological examination. One patient with
increase in the number of women having fertility treatment in the 40s. The moderate endometriosis was diagnosed to have an adenocarcinoma arising
success rates of IVF and IUI are low. HFEA data from 2006 showed that the from endometriotic lesions in the pouch of Douglas. 82.7% (n=105) of
live- rate from IVF in the UK was 11% in the age-group 40-42, 4.6% in the patients had patent tubes, 13.4% (n=17) had unilateral tubal bock and 2.9%
age-group 43-44 and 4% in women over 44. Meta Analysis of (28 RCTs) (n=5) had bilateral tubal block on chromopertubation. 3.1% (n=4) women
showed an increase in clinical pregnancy rate (OR 1.29, 95% CI 1.12 to had unilateral and 2.4% (n=3) had bilateral peritubal adhesions.
1.49) and significantly increased multiple pregnancy rates per woman after
April 19-22, 2009 48

Conclusions: The most common findings in women who had a negative

history, normal examination findings and a normal pelvic ultrasound and
had failed to conceive with super ovulation and IUI was endometriosis.
Though the benefit of treatment for AFS class I (minimal) and II (mild)
endometriosis has been questioned, significant pelvic pathology was seen in
26.8% (n=34) of women. They included moderate endometriosis (AFS 3),
PID or tubal pathology. Use of laparoscopy was of significant benefit, as at
least 1 in 4 women had conditions where treatment was proven to improve
her chances of future fertility. Where resources are limited, it seems logical
to treat patients with normal pelvic imaging for 3 or more cycles of super
ovulation and IUI before proceeding to laparoscopy.
211.7 – EFFECTIVE WAY TO CUT THE POST OPERATIVE ANALGESIC

REQUIREMENT AFTER FEMALE TUBAL STERILISATION.
Kinza Younas, T. Nash, S. Ekladios, T. Abo Kaseem, Ralph Settatree.
Royal College of Obstetrics & Gynaecology London; Sketty Swansea,
Swansea; UK.
Aims and Standards: We have evaluated the effiectiveness of application of
2% lignocaine gel to Filshie clips and the direct infiltration of mesosalpinx
with Bupivacain to relieve postoperative pain as against the control group
where no local analgesic was used. National evidence based guidelines for
male and female sterilisation Jan.2004.
Background: Laparoscopic sterilisation is more painful than diagnostic
laparoscopy probably because of local tissue necrosis and ischaemia at the
site of tubal interruption.
Methods and Materials: 77 healthy women not allergic to local anaesthetic
used had laparoscopic tubal occlusion under general anaesthesia at the
Solihul and Birmingham Heartlands hospitals, UK. Survey forms were filled
in the ward by the nursing staff at 01 hr.post operative to assess pain score
against 10 point numeric rating scale (NRS), at the request of analgesia and
at time of discharge.
Results: Flishie clips covered with 2% lignocaine gel were used in 24/77
while direct infilteration of mesosalpinx in 13/77 cases. 75/77 patients had
skin infiltration at the site of incisions. Pain scores remain similar in 2%
lignocaine gel and Bupivacain group and had significantly longer time to first
analgesia, less pain at 1 hour and shorter recovery time(P value <0.0001) as
compare to the control group. There was no case of failed sterilisation or
adverse reaction to local analgesics.
Recommendation: The application of local anaesthetic gel to Filshie clips is a
safe, non-invasive, and effective method of relieving postoperative pain
during laparoscopic tubal sterilisation as compare to the direct tubal
infilteration which may be associated with haemotoma formation or risk of
bleeding,more time consuming and expertise required for this techni que.
Conclusion: As a safe and quick method Filshie clips soaked with lignocaine
gel should be used to reduce the postoperative analgesic requirement.
More work in the form of large scale audit is needed to support these results
in future.
49 Geneva
April 19-22, 2009 50

ORAL ABSTRACTS – TUESDAY
babies.29 In a review of 165 pregnancies and 200 infants conceived following

300.2 – NEW CHALLENGES FOR OVARIAN-CELL THERAPY AT THE DAWN OF OV, the birth weight and the incidence of congenital anomalies (2.5%) were
THE 21ST CENTURY comparable to those following spontaneous conception or IVF treatment.30 A
novel fertility preservation strategy involves immature oocyte retrieval in an
Alain Gougeon1, Dominique de Ziegler2. Inserm U865, Lyon; 2Gynécologie- unstimulated menstrual cycle31 or from ovarian tissue biopsies,32 followed
Obstétrique II, GH Cochin-St Vincent de Paul, Paris; France by in-vitro maturation (IVM) and OV or EC. IVM has become an effective
Our lecture will provide an overview of the most salient findings that were treatment option for many infertile women,33-37 resulting in the birth of over
recently made in the emerging domain of cell therapy. This will constitute 500 healthy infants38 without any increase in fetal abnormality or
the stronghold on which projections will be drawn for elaborating the miscarriage rates in comparable patients.39, 40 In a pilot study at the MRC on
possible new developments that can reasonably be foreseen for the next IVM-OV, an LBR/C of 20 % per cycle was achieved, including the world’s
decade or so. Expending upon existing data and elaborating from the first four live births from vitrified IVM oocytes.29, 41 Advantages of IVM-OV or
expected future accomplishments to be made in this field, particular EC include: 1) eliminating expensive drugs and monitoring; 2) completing
emphasis will be put on the possible practical applications of ovarian cell treatment within 2 to 10 days;31 3) avoiding hormone-sensitive tumors; and
therapy. In doing so, we’ll primarily focus on depicting how this novel 4) retrieving oocytes at any phase of the menstrual cycle.42 To date, the MRC
approach may help in the future women who suffer from major reproductive has provided fertility preservation to over 130 patients with breast,43 hema-
failure and assist them in their quest for conceiving and becoming pregnant. tological, brain, soft tissue, colorectal and gynecological cancers.
Primary-care physicians and oncologists need to be made aware of the
The reproductive failures at stake that are possible candidate for ovarian cell
fertility preservation options and to provide early discussion with the
therapy include premature menopause, premature ovarian failure (POF)
patients followed by referral, if desired, to an IVF center that offers the full
occurring either spontaneously or triggered by past chemo- and/or radiation
range of fertility preservation options.
therapy for cancers that affected women of reproductive age. In the long
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51 Geneva
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33. Chian RC, Gülekli B, Buckett WM, Tan SL. Priming with human chorionic gonadotropin before
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Engl J Med 1999;341(21):1624-6. adverse effects of controlled ovarian hyperstimulation, suppression of the
34. Chian RC, Buckett WM, Tulandi T, Tan SL. Prospective randomized study of human chorionic
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gonadotrophin priming before immature oocyte retrieval from unstimulated women with
polycystic ovarian syndrome. Hum Reprod 2000;15(1):165-70. during follicle aspiration. Supra-physiological levels of steroids secreted by
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36. Al-Sunaidi M, Tulandi T, Holzer H, Sylvestre C, Chian R-C, Tan SL. Repeated pregnancies and live level. This seems to be the primary cause of the luteal phase defect during
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2008;23:2010-1016. endometrial maturation thus disrupting the delicate mechanism of embryo-
38. Edwards RG. Foreword. In: Tan SL, Chian RC, Buckett WM, eds. In vitro maturation of human endometrial interaction. During COH, estradiol concentrations are
oocytes Basic science to clinical application. London: Informa; 2006:xv-xx.
supraphysiological due to multifollicular maturation. Furthermore,
39. Buckett WM, Chian RC, Holzer H, Dean N, Usher R, Tan SL. Obstetric outcomes and congenital
immediately after ovulation, estradiol concentrations decrease to a greater
abnormalities after in vitro maturation, in vitro fertilization, and intracytoplasmic sperm injection.
Obstet Gynecol 2007;110(4):885-91. extent due to follicular aspiration and early progesterone rise. The decrease
40. Buckett WM, Chian R-C, Dean NL, Sylvestre C, Holzer HEG, Tan SL. Pregnancy loss in level of estradiol is more pronounced due to formation of multiple corpora
pregnancies conceived after in vitro oocyte maturation, conventional in vitro fertilization, and lutea in response to stimulation with HCG from granulosa-lutein cells
intracytoplasmic sperm injection. Fertil Steril 2008;90:546-50. aspirated during oocyte retrieval in agonist or antagonist cycles.
41. Chian RC, Gilbert L, Huang JYJ, et al. Live birth after vitrification of in vitro matured human
oocytes. Fertil Steril:In press. Implantation is a complicated process that involves adequate preparation of
42. Demirtas E, Elizur S, Holzer H, et al. Immature oocyte retrieval in the luteal phase to preserve the endometrium. An important contributor to this preparatory process is
fertility in women with cancer facing imminent gonadotoxic therapy: it is worth a try. Reprod the corpus luteum. The primary function of which is to secrete progesterone
Biomed Online 2008;17(4):520-3.
to induce secretory transformation of the endometrium to facilitate
43. Oktay K, Demirtas E, Son WY, Lostritto K, Chian RC, Tan SL. In vitro maturation of germinal
implantation. During the early stages of its development necessary support
vesicle oocytes recovered after premature luteinizing hormone surge: description of a novel
approach to fertility preservation. Fertil Steril 2008;89(1):228. may be required for the continuation of pregnancy. The corpus luteum
requires continual stimulation by luteinizing hormone (LH) to maintain
adequate production of progesterone which is produced by the action of
301.4 – DIAGNOSIS AND MANAGEMENT OF POOR RESPONDERS human chorionic gonadotropin (HCG) on the LH receptor during pregnancy.
Marc Germond, Centre de Procréation Médicalement Assistée (CPMA) and Stimulated IVF cycles are associated with luteal phase defect. In order to
FABER Foundation, Lausanne, Switzerland. overcome this, different doses, durations and types of luteal phase support
The definition of a poor response to ovarian stimulation varies according to (LPS) have been evaluated. There is still no agreement regarding the optimal
whether an Ovulation Enhancement (OE) prior to timed sexual intercourse or a supplementation scheme.
Controlled Ovarian Stimulation (COS) for oocyte recovery and in vitro HCG is a time honored hormone that has been used for luteal phase
fertilisation is considered. As a result, there is no universal definition. Most support. Due to the increased risk of hyperstimulation, it has largely been
often, an absence of follicular development is considered as a poor response replaced by progesterone.
in the first case, and the recovery of less than 5 metaphase II oocytes in the
second one. Progesterone can be administered orally, vaginally, or intramuscularly. It
leads to higher serum progesterone levels and better pregnancy rates.
The poor response can have a genetic, immunological, iatrogenic or
biochemical origin. In most cases, the age of the patient is implicated in this Dydrogesterone (DG) was introduced to support the luteal phase of
phenomenon, which would remain undiagnosed if no treatment were stimulated IVF cycles. DG, a retroprogesterone is a biologically active
undertaken. metabolite of progesterone. Its anti-estrogenic effect on the endometrium
helps in achieving the desired secretory transformation.
Echographic and hormonal diagnostic tools have been evaluated in
numerous studies. A consensus tends to involve the ovarian volume and the The addition of estradiol (E2) seems to be beneficial in long GnRH agonist
number of antral follicles evaluated by ultrasound, an AMH level of less than protocol but not in the short GnRH agonist and GnRH antagonist protocol.
3 IU/l, a FSH >12 IU/l, as well as a low E2 on the 3rd day of the cycle; The randomized trials showed a beneficial effect of use of estrogen in terms
however, no ideal combination of these factors could be established. A of pregnancy rates (1). However, the latest meta-analysis published in 2008
stimulation trial constitutes the ultimate step able to confirm the clinical could not reveal any beneficial effect of E2 co-administration in stimulated
prognosis. IVF cycles.
The literature agrees on a maximum dose of 450 IU that can be Several other agents have been mainly used as adjuncts to progesterone.
administered per day. The time at which treatment can be started (luteal or These are HCG, estradiol, GnRH agonists, aspirin, and various others.
follicular phase), the concomitant administration of LH, GnRH agonists or Estrogen has been advocated as an adjuvant to progesterone for LPS.
antagonists are still the subjects of many publications. Estrogen can be administered either orally or transdermally. While studies
failed to corroborate the effective results.
We report on our experience based on 3’561 oocyte recoveries performed
on patients aged 35 to 45 years. The results were analysed retrospectively in Several adjutants together with mainly progesterone have been administered
respect to the oocyte and zygote numbers, as well as to the implantation during the luteal phase with the aim to increase the implantation rate. The
and delivery rates. These data were related to the number of FSH units addition of ascorbic acid or prednisolone has not been found to be beneficial
administered and re-evaluated in the light of the patient’s age. (2, 3).
Our conclusions confirm that the therapeutic response remains the best Aspirin has been advocated both to increase ovarian responsiveness and
April 19-22, 2009 52

implantation. Although a very recent meta-analysis of the prospective transvaginal ultrasound guided follicle aspiration was introduced, which
randomized studies showed that aspirin did not increase pregnancy and simplified the retrieval and made it more effective. .In 1992 the introduction of
delivery rates in the ART setting (4). ICSI improved dramatically the fertilizations rate. Assisted hatching, the
Despite the widely adopted practice of luteal phase support especially in availability of new transfer media and blastocyst stage transfer also improved the
ART cycles there is still the need for properly designed and adequately implantations rate significantly.. Therefore fewer follicles and oocytes were
powered randomized studies to determine the agents/s that are associated enough to achieve success in IVF treatment. This brought up the issue of mild
with higher implantation rates. stimulation again (R.G. Edwards 1996). The discovery and application the GnRH
antagonists enhanced the use of the milder stimulation procedures. Due to the
References: successful application of ICSI in the treatment of the male infertility, the number
Farhi J, Weissman A, Steinfeld Z, Shorer M, Nahum H, Levran D. Estradiol of the healthy and young IVF patients is increasing. The single embryo transfer-to
supplementation during the luteal phase may improve the pregnancy rate in avoid the multiple pregnancies- is also more widely used. Because of these
patients undergoing in vitro fertilization-embryo transfer cycles. Fertil Steril, facts, mild stimulation in the current IVF practice has an increasing importance.
2000; 73: 761-6. Clinical research has to focus on this kind of stimulation, which is safe, effective
and cost saving.
Griesinger G, Franke K, Kinast C, Kutzelnigg A, Riedinger S, Kulin S Kaali
SG, and Feichtinger W. Ascorbic acid supplement during luteal phase in IVF.
J Assist Reprod Genet, 2002; 19: 164-8. 303.2 – OUTCOME OF THE RECENT IVF PROGRAM TRANSFERRING SINGLE
Ubaldi F, Rienzi L, Aniballo S, Iacobelli M, Cobellis L, Greco E. Low dose BLASTOCYST RETRIEVED ON MINIMAL STIMULATION CYCLE AND
prednisolone administration in routine ICSI patients does not improve FROZEN BY VITRIFICATION METHOD
pregnancy and implantation rates. Hum Reprod, 2002; 17: 1544-1547. Keiichi Kato, Kato Ladies Clinic, Tokyo, Japan
Gelbaya T, Kyrgiou M, Stern C, Nardo L. Low dose aspirin for in vitro Introduction: One of the major concerns of the modern assisted reproductive
fertilization: a systematic review and meta-analysis. Hum Reprod Update, technology (ART) is to reduce the number of oocytes retrieved and transferred
2007; 13: 357-364. keeping pregnancy rate high. Ovarian hyperstimulation impose physical and
financial burden on patients and transfer of multiple embryos cause multiple
pregnancy, which cause maternal complications and neonatal morbidity and
302.2 – CRYOPRESERVATION OF OOCYTES
mortality. To overcome these issues, we have implemented a new IVF program in
Ri-Cheng Chian, McGill Reproductive Center, Department of Obstetrics and which small number of oocytes were retrieved under the clomiphene-based
Gynecology, McGill University, Montreal, Canada. minimal stimulation cycle and frozen by vitrification method, and then followed by
Improving success rate associated with the use of cryopreserved human single blastocyst transfer under hormone-controlled or un-stimulated natural
oocytes would be an important advance in human assisted reproductive cycle. The purpose of this presentation is to report the most recent results of the
technology (ART). The objectives of this presentation are to evaluate the program in which more than 10,000 ET cycles were performed in 2007.
survival rates and pregnancies of cryopreserved human oocytes following Materials and Methods: In 2007, 19,305 IVF cycles of minimal stimulation and
vitrification. As a clinical trial, a total of 38 women were included in the 10,115 cycles of embryo transfer were performed. All cycles that the IVF protocol
study, in which women underwent ovarian stimulation, their mature and specified below were completed included in this study. The mean age of patient
immature oocytes collected and vitrified. The mean age of women was was 39.4±4.8 years old. There was no exclusion criterion on cause of infertility.
31.5±0.5 years. The vitrified oocytes were thawed and inseminated using
Oocytes were retrieved on the minimal stimulation cycle: clomiphene citrate (50-
intracytoplasmic sperm injection (ICSI), and the resulting embryos were
100 mg/day) was administered from day 3 up to one days before the oocyte
transferred in a subsequent menstrual cycle. The Hospital Research Ethics
retrieval with or without hMG or rFSH (50-150 IU/every other day, from day 8 or
Board (REB) has approved the protocol. The patients were given fully
later), to elaborate 1-4 follicles. LH surge was induced by nasal GnRH agonist
explanation for the procedures and a written consent forms were obtained.
(Sprecure®, 300 μg) when the size of the leading follicle is more than 21-
As the procedure of vitrification, the oocytes were suspended in
24mm). 34-36 hr later, oocytes were retrieved and inseminated conventionally or
equilibration medium (7.5% ethylene glycol and 1,2-propanediol) for 5 min,
by ICSI cultured to blastocysts. Fertilized oocytes were cultured individually in 20
and then the oocytes were transferred to vitrification medium (15% ethylene
μl of Cleavage medium (SAGE, USA) from day 1 to day 3, and then in Blastocyst
glycol and 1,2-propanediol + 0.5 M sucrose) for 45-60 seconds at room
medium (SAGE) from day 4 to day 6. In 4449 cycles, embryo (mostly day 5
temperature. The oocytes were loaded on a special designed device, McGill
blastocysts) were frozen by vitrification method using Cryotop® as described
Cryoleaf, and immediately plunged it into liquid nitrogen for storage. For
elsewhere. Single ET was performed using thawed embryos on the next cycle or
thawing, the McGill Cryoleaf was directly inserted into 37ºC thawing medium
later (4,449 cycles), or using fresh embryo (mostly day2 embryo) (5,666 cycles).
(1.0 M sucrose) for 1 minute. The thawed oocytes were transferred to 0.5 M
For frozen-thaw transfer, uses of hormonal replacement cycle were a choice.
and 0.25 M sucrose medium for 3 minutes respectively, and then washed
twice with culture medium for insemination using intracytoplasmic sperm Results: The average number of oocyte picked up per cycle was 1.35 (26,100 /
injection (ICSI) method. As the results, a total of 463 oocytes were vitrified 19,305 cycles). At least one blastocyst was achieved in 54.6% of 19305 cycles.
from these 38 women. Of those, 383 (82.7%) oocytes were survived post- Average number of ET performed was 1.01. Overall survival rate of frozen
thaw. Following insemination by ICSI, 287 oocytes were fertilized normally blastocyst was 95.0%., comparable to that of cleavage stage (98%).
(75.0%), and 17 patients became pregnant (44.7%) after transferring the The pregnancy rates achieved frozen-thawed single blastocyst transfer were
resulted embryos (3.7±1.1). Implantation rate was 18.1% (24/133). 41.3% (per ET), which were significantly higher than that obtained by fresh day2-
Obstetric and perinatal analysis of pregnancies and live births indicates there embryo transfer (19.8%). Pregnancy rate showed age-dependent decrease in
is no adverse outcome associated with oocyte vitrification. This result both frozen-thawed ET as in fresh ET, the positive impact of frozen ET was
indicates that human oocytes can be cryopreserved successfully with the enhanced in the older age group (≥35 y.o.) compared to the younger group (<34
procedure of vitrification and that vitrification method may be efficiently y.o.).
used for a program of cryopreservation of human oocytes in ART. Oocyte
vitrification technology may offer to women for fertility preservation. The overall multiple pregnancy rates were as low as 1.3%, almost comparable to
that of normal Japanese population. Ectopic pregnancy rate was 0.3%.
Conclusions: The clomiphene-based minimal stimulation IVF programs gave
303.1 – THE ROLE OF THE MILD STIMULATION IN THE CURRENT IVF satisfactory outcomes, in terms of pregnancy rate, comparable to that achieved
PRACTICE by standard controlled ovarian hyperstimulation methods in Japan, patient
Artur Bernard, Kaali Institute, IVF Center, Budapest Hungary. compliance and singleton pregnancy. Use of blastocyst ET also reduced ectopic
pregnancy rate to the level of non-IVF pregnancy.
The birth of the first IVF child was achieved from a natural cycle./Steptoe and
Edwards 1978/. The aim of the ovarian stimulation has been to produce, retrieve
and fertilize more oocytes, and select afterwards the best embryos for transfer.
Therefore in the 80’s most centers used high dose stimulation to increase
oocyte/embryo yield, and therefore to improve pregnancy rates. In 1991
53 Geneva
Conclusions: Although conventional procedure in mechanical infertility patients

303.3 – CONTROLLED OVARIAN STIMULATION, MINIMAL STIMULATION OR NO is still superior, IVM was found to be an efficient method in patients with PCO
STIMULATION FOR IN VITRO FERTILIZATION: IS THERE AN OPTIMAL syndrome. Since our results are based on a small number of patients, further
STIMULATION? studies are still ongoing.
John Zhang, Osamu Kato. New Hope Fertility Center, New York NY, USA and Kato
Ladies Clinic, Tokyo, Japan. 304.2 – OXYGEN CONSUMPTION BY SCANNING ELECTROCHEMICAL
The success of IVF treatment is assessed by four criteria consisting of pregnancy MICROSCOPY (SECM) FOR HUMAN IVM COC & EMBRYO WITH PCO
outcome, safety, patient comfortableness and cost. With improvement of embryo PATIENTS
preservation with vitirfication and promotion of single embryo transfer it is the Hiroaki Yoshida, Yoshida Ladies Clinic, Reproductive Research Center, Sendai,
time to reassess various stimulation protocols in terms of safety and efficiency. In Japan
United States of American almost 95% of IVF cycles are performed through daily Introduction: Recently, in vitro maturation (IVM) has been used for human
gonodotropine injection with GnRH analogs or antagonists to control premature assisted reproductive technology (ART) for women with PCO. IVM-IVF was
ovulation (conventional IVF). Since 2004 we have elected to use primarily a mild performed for patients with PCOS in 2008. Among a total of 186 retrieved
stimulation protocol with clomiphene citrate (CC) in combination with a small oocytes, the maturation rate was 60.7%, the pregnancy rate was 29.4%. As the
addition of human menopausal gonadotropin (hMG) in our IVF cycles (minimal both maturation and pregnancy rates were considered to be unsatisfactory low,
stimulation IVF). the simultaneous use of nuclear and cytoplasmic maturation systems was
Clomiphene citrate (50 mg) was initiated orally each day, beginning on Day 3. considered in order to clarify the relationship between cytoplasmic maturation
Subcutaneous administration of 75–150 IU of hMG every 48 hours was begun on and mitochondrial distribution or function. Furthermore the mitochondrial
Day 5 or 8 depending on Day 3 FSH level. For patients with Day 3 FSH level is function of IVM oocytes and the mitochondrial membrane potential were
more than 15 no Clomiphene citrate will be give and patient will only under observed using transmission electro microscopy (TEM) and scanning
natural cycle IVF. A gonadotropin releasing hormone agonist (GnRHa) nasal spray electrochemical microscopy (SECM). As oxygen consumption is an ideal indicator
(Synarel) was administered to trigger an endogenous LH surge for final of metabolic activity. SECM measuring systems have been demonstrated to
maturation of the oocytes. Oocyte retrieval was performed 34 hours after the successfully non-invasively the measure respiration activity of single embryos
administration of the spray from several species. It has been reported that the bovine embryos with higher
In this presentation we wish to compare various protocols of ovarian stimulation oxygen consumption are better candidates for further development of good
and would like to propose a concept of optimal ovarian stimulation which is quality embryos and yielded higher pregnancy rates. Despite this apparent
based on daily FSH levels. Traditionally daily dosing of stimulation is tailored to correlation between respiration activity and embryo quality, .oxygen
patient age, ovarian reserve and how follicular development and estrogen consumption in human IVM oocytes and embryos has yet to be evaluated.
hormone rising. A FSH level based stimulation will provide an opportunity to give Materials and Methods: Cumulus oocyte complexes (COC) and oocytes were
the least medicine need to generate certain number eggs in each IVF cycles to transferred into a plate filled with HFF99 medium. A microelectrode was used to
minimizing hyperstimlulation and save the cost of medications. scan along the z-axis from the edge of the sample and the oxygen consumption
rate was calculated based on the spherical diffusion theory using custom
software. Measurements of each oocyte were performed rapidly (within 1 min).
304.1 – THE BENEFIT OF IVM IN PCOS PATIENTS Subsequently, a sample of each oocyte was collected for observation by TEM.
Yona Barak, Olga Sikorska, Tomasz Rokicki, Aneta Karwacka, Tomasz Dworniak, Results: We classified morphologically the COC and oocyte size from grade1 to
Ricardo Faundez. InviMed, European Centre of Motherhood, Warsaw, Poland. 5. Our observation suggested that. Oocyte maturation rate was decreased from
Objective: In vitro maturation (IVM) was established as a new method for treating grade 1 to 5. The consumption for a single oocyte was as follows: GV 0.49, MI
infertility. The procedure was applied in our centers in patients showing 0.47, MII 0.41 Fx1014/mols-1. Weak correlation with COC size and oxygen
Polycystic Ovary Syndrome (PCOS). consumption rate before and after 26 hrs culture was observed; however there
was no significant difference between oocytes before and after culture. These
Design. Rate of maturation, fertilization, embryo scoring and pregnancies were
findings suggested that the oxygen consumption rate is representative of the
compared in patients with PCOS, undergoing IVM procedure (study group) with
COC respiration activity and that it can be used to indicate adequate oxygen
patients undergoing IVF due to female mechanical cause of infertility (control
consumption. There were no significant difference in the mean of oxygen
gruop).
consumption at each cleavage in embryos (0.2 ~0.56×1014/mols-1).
Materials and Methods: Twenty nine patients determined as PCOS underwent
Conclusion: The COC mitochondria which were enlarged and showed elongated
32 IVM cycles. The control group (n=28) underwent conventional ICSI procedure.
morphology. Good quality oocytes have rich mitochondria and have gap junction
IVM patients were mildly stimulated using 150 I.U. Gonal-F for 3-4 days. IVM was
between cumulus cells. IVM COC was found to be important for oocyte
performed according to the procedure of MediCult IVM System (MediCult,
maturation and embryo development and it is strongly related to oocytes via gap
Denmark). ICSI was performed 30 hours (Day 0) after IVM medium (MediCult,
junctions. There was no significant difference embryo consumption which
Denmark). Injected oocytes were then cultured in MediCult ISM1 medium.
moderate respiration rate of embryos showed high developmental rate to the
Patients of control group were stimulated in the short protocol using 75 IU of
blast cyst. Further study of cytoplasmic maturation and mitochondrial function
Fostimon with GnRH antagonist Cetrotide. Ovulation was induced by Ovitrelle.
from IVM oocytes and embryos is required.
Following ICSI, oocytes cultured identically as PCOS oocytes.
Number of retrieved oocytes, rates of fertilization, cleavage, quality of embryos
and pregnancies were compared with PCOS and control group, by Yate’s 304.3 – CLINICAL DATA OF IVM FOR PCO PATIENTS
corrected 2 test. Results. Mean number of aspirated cumulus enclosed oocytes Yoshiharu Morimoto, The Centre of Reproductive Medicine and Infertility, IVF
(OCCs) per patient was 8,8 and was significantly lower in comparison to the Japan, Osaka, Japan.
control (17,0). A significant difference in the rate of OCCs which reached the MII
stage was observed after 30h in the PCO group in comparison to the control Polycystic ovary syndrome (PCO) is a disease including ovulation dysfunction of
(138/257 (53,7%) vs 409/509 (79,4%), respectively, p<0,01). No differences ovaries or amenorrhea, hirsutism and obesity. It is one of the most important
were observed in fertilization rate (122/156 (78,2%) vs 337/377 (89,4%) origins of infertility and is difficult to be treated. In the treatments of PCO,
however a significant difference was found in cleavage rates (101/119 (84,9%) ovulation induction, birth control pills, operative procedures such as wedge
vs 177/184 (96,2%), p<0.05). A higher value of the mean fragmentation grade of resection of ovary and ovarian drilling under laparoscopy have been applied.
4-5cell stage embryos was observed on day 2 in the PCO group in comparison to Recently insulin sensitizing medications such as Metformin, Piogritazone and
the control group (1,6 vs 1,3 respectively; p<0.01). Nine pregnancies out of 26 Rosiglitazone have been added as an option.
embryo transfers (ETs; 34,6%) were achieved in the PCO group; 23 ETs In Vitro Maturation (IVM) has been first reported by Cha et al. (1991) as one of
performed in the control group resulted in 13 pregnancies (46,4%; mean of 2.3 treatments for PCO and nowadays spread worldwide. In the initial stage, the
vs 2.0 embryos per patient, respectively). clinical success rate of IVM was low, however, it has been improved remarkably
by the development of media specified for IVM and puncture needle. Hence, IVM
April 19-22, 2009 54

may be a preferable option to be selected in terms of prevention of ovarian condition are caused by abnormal pituitary function; abnormal steroidogeneis in
hyperstimulation syndrome (OHSS)and accuracy. ovary and adrenal gland; and abnormal tissue respond to various hormones. Due
We had the first success in IVM procedure in 1999 in Japan. The first baby was to the complexity of the disorder, the management strategies should be designed
born next year, and first baby by frozen embryo transfer by IVM was born in to tailor individual patient. Based on each patient’s presentation and the goal of
2001. seeking for medical care, we should form a short and long term treatment plan.
This presentation will address the current management strategies on androgen
Our indication for IVM procedure is mainly for PCO. But we apply this procedure excess; ovulatory dysfunction; insulin resistance; endometrial protection; obesity
for the patients with normal menstrual cycles, rarely for the cases of poor quality and metabolic syndrome. Various options in ovulation induction and ART in
embryo. management of PCOS will also be discussed.
We start follicle monitoring from Day 7 of the menstrual cycle. At the day we can
recognize at least two follicles of the diameter of over 7mm, immature oocytes
are collected. The program should be cancelled when we see a dominant follicle 306.3 – HUMAN OOCYTE ULTRASTRUCTURE IN IVM
or a ovarian cyst. HCG priming by 10000 iu is done 36 hours before oocyte Yoshiharu Morimoto, IVF Namba Clinic, The Centre for Reproductive
retrieval. Follicles are aspirated by IVF OSAKA IVM needle (Kitazato Biopharma Medicine and Infertility, Osaka, Japan.
Co.ltd, Tokyo, Japan). The IVF OSAKA IVM needle is designed by ourselves and is Introduction: In vitro maturation technique (IVM) has been introduced world
composed of two segments of puncture needle and holding needle. MediCult IVM wide. In order to improve the technique toward better result, the
medium is arranged with 10% patient’s serum and HCG and FSH. Retrieved understanding of the oocyte maturation, especially of cytoplasm, is
oocytes are cultured for maturation for 24 hours and all matured oocytes are important. The ultrastructural investigation for oocyte in IVM may open the
inseminated by ICSI. black box of maturation mechanism.
Up to 2007, we had 705 IVM cases and 91 pregnancies. The live birth and on- Oocyte maturation: Human primordial germ cells in the fourth week of
going pregnancy rate was 78% and the rate of abortion and stillbirth was 23.1%. embryonic development migrate to the coelomic epithelium of the gonadal
We had only one case of malformation which was Goldenhar syndrome. ridges and develop to oogonia. Oogonia forms and exists as a manner of
From these data, we strongly recommend to select IVM option for the treatments nest. The first follicles are recruited by the action of neurotrophic factors
of PCO patients as a first choice. such as brain derived neurotrophic factor (BDNF), nerve growth factor and
neurotrophins(NT). On the way of its process, one layer of follicular cells
covers oocyte and follicle development comments. For the transformation,
304.4 – USE OF LETROZOLE IN POLYCYSTIC OVARY SYNDROME (PCOS) Kit system and anti-mullerian hormone has important roles on the process.
Makio Shozu, Tomoya Segawa2, Satoshi Kawachiya2, Osamu Kato2. Department of Oocyte maturation process is divided into several categories, such as
Reproductive Medicine, Graduate School of Medicine, Chiba University, Chiba; nuclear, cytoplasmic, zona pellucida and follicular cell maturation. In order
2
Kato Ladies Clinic; Shinjuku, Tokyo; Japan. to acquire the competence for fertilization and further development,
An aromatase inhibitor, letrozole, has recently been introduced into variable accomplishment and harmony between each factor are needed.
situations of ovarian stimulation. The major application of the drug is Nuclear maturation: Meiosis is a reducing process of chromosomes from
superovulation in IVF or IUI protocols, in which letrozole is administrated diploid to haploid in gametes. At the time of migration to the coelomic
combined with gonadotropins to increase the number of oocytes ovulated and the epithelium of the gonadal ridges, primordial germ cell undergoes mitotic
resulting pregnancy rate. Letrozole is also used for PCOS in an attempt to achieve division. The first meiotic division is a transition from oogonia to oocytes.
single ovulation and singleton pregnancy. This is because, in contrast to The completion of nuclear maturation occurs with germinal vesicle
clomiphene citrate (CC), letrozole potentially allows natural selection of a breakdown by the action of maturation promoting factor and finishes at
dominant follicle. An additional advantage of theoretical interest is seen with extrusion of polar body.
letrozole use for PCOS compared to CC. The shorter half-life of letrozole favors
rapid recovery of endometrial thickening, which may enhance implantation rate. Cytoplasmic maturation: Cytoplasmic maturation of oocyte may be an
Ovarian hyperstimulation syndrome may also be avoided by reducing the number essential phenomenon for the oocyte to acquire competence to fertilize and
of oocytes. These advantages of letrozole over CC have been postulated based on cleave. Oocyte undergoes its maturation in harmonization with maturation of
clinical studies in infertile women with heterogenous etiologies, including PCOS, nucleus and cytoplasm. A plenty of mRNA including maternal genes are
and the advantages in PCOS remain inconclusive. The effectiveness of letrozole in accumulated in the cytoplasm. The fact is shown as increase and
infertile patients with PCOS (2003 ESHRE/ASRM criteria) was thus assessed. Over differentiation of organella.
400 PCOS patients treated with or without letrozole were included and reviewed Ultrastructure of the ooplasm in maturing process: Immature and matured
for this analysis. With concomitant use of FSH (75 or 150 IU/every other day, days oocytes were donated by PCO patients aged from 27 to 33 years who
8 - 12), letrozole (2.5 mg/day, days 3 - 7) was as successful for ovulation underwent IVM program in the two centers of infertility treatments in Osaka
induction as CC (100 mg/day, days 3 - 7), with ovulatory rates of 89% and 83%, after informed consent. Oocytes were cultured in IVM medium NG 1.3 (
respectively. Total dose of FSH was significantly lower in the letrozole+FSH group Medicult, Denmark), 10% patient serum, 100 IU/L human chorionic
(mean, 242 IU; range, 75 - 1050 IU) compared to the letrozole+CC group (mean, gonadotropin and 75 IU/L follicle stimulating hormone under the
406 IU; range, 100 - 1200 IU). Single ovulation rate was significantly higher in the atmosphere of 5% CO2, 5% O2 and 90% N2. Those oocytes were fixed and
letrozole+FSH group (68%) compared in the CC+FSH group (38%). Clinical ultrathin sectioned and observed by electron microscope at 0, 6, 12 and 24
pregnancy rate tended to be higher in the letrozole+FSH group (44%) than in the hours of culture.
CC+FSH group (34%), although this difference was not significant. Letrozole is Immature oocyte which is still in GV stage cultured in vitro for 6 hours
thus as effective as CC in terms of ovulation induction and may be superior to CC showed that granulosa cells surrounding oocyte looked very active and
in terms of single ovulation even when administered with FSH. included a plenty of lipid droplet. Mitochondria were dispersed in cytoplasm
and cortical granules were prominent on the edge of the plasmalemma.
304.5 – PCOS: MANAGEMENT STRATEGIES IN 2009 After 24 hours of in vitro culture, the ultrastructure showed remarkable
change. In the nucleus, germinal vesicle was broken down and cortical
Frank Yelian, LIFE Clinic, Los Angeles, CA, USA. granules disappeared on the edge of the cell. More noteworthy character is
Polycystic ovarian syndrome is one of the most common endocrine disorders in that of mitochondria. They have migrated over into the center of the cell,
reproductive women. It is often presents with oligomenorrhea, androgen excess, increased in number and aggregated. Furthermore microvilli on the surface
insulin resistance, and obesity. The short and long term consequences of PCOS of the oocyte showed morphological remarkable development.
include ovulatory dysfunction and infertility; metabolic syndrome and type II Conclusion: Despite during the short period of 24 hours, the cell
diabetes; endometrial hyperplasia and cancer; dyslipidemia and cardiovascular construction has dramatically changed in its ultrastructure. The remarkable
diseases. PCOS is now considered to be a complex genetic disorder. It has been development of microvilli on the surface of the oocyte indicates the active
suggested that the disorder is a result of one of many intrinsic variant genetic communication from the inside to the outside through plasma membrane.
traits that interact with other congenital or environmental factors to cause the The purpose of the oocyte maturation is defined as preparation for
endocrine dysfunction. The pathophysiology and clinical symptoms of the fertilization and cleavage. The movement and aggregation of the activated
55 Geneva
mitochondria indicates the competence acquirement in preparation for (Kuwayama 2005) was used to vitrify the oocytes. The oocytes were equilibrated
coming big events by charging up cell activity. in 7.5% ethylene glycol and 7.5% DMSO in modified medium 199 (M-199) for
Those characteristic changes in ultrastructure may be a part of reality of 15min before being transferred into the vitrification solution (VS) for 30 sec.
oocyte cytoplasmic maturation. Oocytes were then transferred into onto the cryotop with minimum volume, and
immediately submerged into liquid nitrogen.
Results: Oocyte retrieval and cryopreservation was successful in 89% (54/61) of
307.1 – FERTILITY PRESERVING OPTIONS OF REPRODUCTIVE AGE CANCER the patients. No patients had any adverse side effects using the minimal ovarian
PATIENTS stimulation protocol and aspiration with a 22 gauge needle under local
Peter Kovacs, Steven G. Kaali. Kaali Institute IVF Center, Budapest, Hungary. anesthesia. The mean age of the patients was 26.0 (±4.8, S.D.). The mean
numbers of oocyte retrieval cycles per patient was 1.79, and the mean number of
Over the past few years the number of reproductive age survivors of cancer
retrieved oocytes per patient was 7.7, and per cycle was 4.3. The mean number
treatment has increased as a result of newer surgical methods and improved
of morphologically normal cryopreserved oocytes per patient was 6.4, and per
chemotherapeutic and radiation therapy regimens. Survival rates of hematologic
cycle was 3.5. The type of cancers included acute and chronic leukemia,
malignancies (most lymphomas, leukemias are diagnosed in patients under the
malignant lymphoma, aplastic anemia, and myelodysplastic syndrome.
age of 40) have increased to 50-90%. Ten to forty percent of gynecologic
malignancies are also diagnosed in reproductive age women. Less destructive Conclusion: Our data showed that the minimal ovarian stimulation protocol using
surgical options, a wider availability of medical options and improved assisted CC for oocyte retrieval with a 22 gauge needle and with local anesthesia was a
reproductive technology all offer hope to a significant proportion of these women. safe, simple, effective method of preserving fertility for unmarried cancer
patients.
Because of these developments the fertility concerns of these women have to be
addressed. As most treatment methods will still severely compromise or even
destroy ovarian function the pre cancer treatment and post cancer treatment 307.3 – SUCCESSFUL VITRIFICATION OF HUMAN OOCYTES
options need to be reviewed with the patient when appropriate. There are
surgical, medical and ART options that may be utilized to maintain fertility. Masashige Kuwayama, Advanced Medical Research Institute of Reproduction,
Kato Ladies’ Clinic, Tokyo, Japan.
Surgical options should be considered for patients diagnosed with gynecologic
malignancies. They include conservative surgery for certain type and stage Recent drastic advances in cryobiology have made it possible to preserve various
ovarian cancers, radical trachelectomy for early stages of cervical cancer and types of reproductive cells with relatively little loss of viability. Ultra-rapid
adnexal lateral position in case abdominal radiation is planned. vitrification, the alternative cryopreservation method seems to be a powerful tool
to any biological specimens, which cannot be preserved by the conventional slow
Medical options (GnRH agonist use) may help to reduce the toxic effects of freezing and previous vitrification. Ultra-rapid vitrification realized the successful
chemotherapy. Progestin therapy is an alternative to surgery when well- clinical use of vitrification not only for human PN zygotes, cleavage stage
differentiated, localized, early stage endometrial cancer is diagnosed. embryos and blastocysts but also for oocytes.
ART is playing an increasingly important role in the management of these The purpose of this lecture is first to introduce the history of vitrification of
patients. For a long time embryo cryopreservation meant the only option. While it human IVF, and the mechanism of vitrification of the cells, how to maintain the
is still considered the most effective technique it cannot be used in all cases. In high viability of the oocytes and embryos under liquid nitrogen temperature, for
recent years oocyte freezing and ovarian tissue cryopreservation offer further your basic knowledge.
options.
To provide evidence for the potential significance of vitrification, achievements
Finally, alternative options like donor egg use or adoption need to be mentioned with the Cryotop technology, an advanced version of the “minimal volume
to complete the list of choices these patients have. approaches” is analyzed. This technology alone has resulted in more healthy
During the presentation the role of the above-mentioned surgical, medical, ART babies after cryopreservation of any stages of embryos than any other
methods will be reviewed including our own cases as examples. We wish to cryopreservation technique, and more successful human oocyte vitrification
stress the importance of appropriate counseling of these patients which often resulting in normal births than any other cryopreservation method. The value of
requires a multidisciplinary approach including, psychiatrists, internists, this method is also demonstrated by achievements in the field of domestic
surgeons, gynecologist, oncologists and fertility experts to address the problem. animal embryology.
Cryotop method has been applied to more than 100,000 clinical cases of oocytes
and embryo cryopreservation for these 8 years, and is now used in more than
307.2 – THE CURRENT APPROACH TO OOCYTES VITRIFICATION FOR CANCER
800 IVF faculties in 12 countries producing excellent clinical results (nearly 100
PATIENTS IN JAPAN
% of post-thaw survival rates for PN zygotes, 4-cells stage embryos, blastocysts
Takafumi Utsunomiya1, Osamu Kato2, Hirofumi Kamiya3, Hiroaki Yoshida4, Yasuhito and oocytes) especially the method actually realized the oocytes cryopreservation
Michikura5, Masanori Ochi6, Yuji Fujino7, Kohji Yano8, Yasuyuki Mio9, Shokichi as an effective protocol in Human IVF. More than 450 of healthy babies have been
Teramoto10. 1St Luke Clinic, Oita; 2Kato Ladies Clinic, Tokyo; 3Kamiya Ladies already delivered from Cryotop oocytes vitrification.
Clinic, Sapporo; 4Yoshida Ladies Clinic, Sendai; 5Towako Ladies’ Clinic, Komatsu;
6 Human oocytes banks for unmarried young cancer patients and for egg donation
Yume Clinic Nagoya, Nagoya; 7Fujino Ladies Clinic, Osaka; 8Yano Maternity
program have also established and been giving dream and courage of life for the
Clinic, Matsuyama; 9Mio Fertility Clinic, Yonago, 10Shinbashi Yume Clinic, Tokyo;
patients to improve their quality of lives as women.
Japan.
Objective: Aggressive chemotherapy and radiotherapy have greatly enhanced
the life expectancy of young cancer patients, but these treatments cause massive 307.4 – CRYOPRESERVATION OF PREPUBERTAL TESTICULAR TISSUE
destruction of the ovarian reserve resulting in infertility or sterility. However, Atsumi Yoshida, Yoko Kurotaki, Miho Tanaka, Hiroki Suzuki, Kiba Park Clinic, Kiba,
oocyte cryopreservation can preserve their fertility of these patients after cancer Koto-Ku, Tokyo, Japan.
treatment. We applied a minimal ovarian stimulation protocol using clomiphene
citrate (CC) to retrieve mature oocytes, and the cryotop vitrification method to For the progress of recent advances in cancer treatments, cure rates of childhood
cryopreserve them in Japan. cancers are very high. However, these treatments may prove toxic to the testis.
Fertility preservation is becoming an important issue in the management of the
Materials and Methods: Thirty four unmarried hematopoietic defect patients quality of life of prepubertal boys undergoing gonadotoxic treatment. As these
with informed consent who underwent the CC cycle from January, 2007 to patients do not yet produce spermatozoa for freezing, the only theoretical option
October, 2008. Fifty mg CC was administered from cycle day 3 and 75 IU for preservation of fertility in these boys is the preservation of the spermatogonial
recombinant FSH was administered every other day from day 8 until the leading stem cells for autologous intratesticular stem cell transplantation or in vitro
follicle developed to 18 mm in diameter. Administration of CC was then stopped, culture of spermatogonia. Currently, spermatogonial stem cell transplantation is
and 300μg GnRH-agonist (buserelin) was given as a maturation trigger. Oocytes considered to be the most promising tool for fertility restoration in young cancer
were retrieved from 30 to 36 hrs following the administration of the GnRH- patients. The spermatogonial stem cells are the male germline stem cells. They
agonist using a 22 gauge needle with local anesthesia (Teramoto, 2007). The can self renew to maintain the stem cell population and produce large numbers
retrieved oocytes were denuded before vitrification. The cryotop method of differentiating cells of the spermatogenic line. Spermatogonial stem cell
April 19-22, 2009 56

transplantation was firstly introduced in the mouse by Brinster et al. In animals, axis of the microscope. The light axis and focus distance were appropriately fine-
the initial method for transplantation was intratubular microinjection in the tuned and it was possible to firmly fix the lenses placed at the front with an
seminiferous tubules. In human, the most common infusion technique for germ additional flexible arm for the surgical microscope. Magnification was improved
cells is ultrasound guided injection into the rete testis. Most studies on by attaching a contact-type lens flexible arm for ophthalmology use to a surgical
cryopreservation of testicular tissue aimed at preserving sperms for microscope. In addition, it was possible to clearly observe inner structure of the
intracytoplasmic sperm injection. Testicular sperm freezing is carried out using in testicular tube by obtaining an appropriate lighting angle and height with an
many instances. The simple rapid method of suspending straws in the external light source (fiber-type). To fix the operational distance, “the direct
uncirculated liquid nitrogen, whereas in freezing of spermatogonial stem cells, it contact method” was employed, and three-dimensional views were obtained by
is necessary to accomplish freezing by lowering slowly the temperature by binocular vision. (2) By attaching ophthalmology lenses as an adaptor,
means of a programmed freezer as is the case with round spermatids and late magnification and stability of the attachment were improved. (3) An appropriate
spermatids. At this institution, we make it a rule at present to perform freezing of lighting angle and proper luminous intensity were obtained with an external fiber
testicular tissues in a programmed freezer, using a cryoprotectant solution light source.
containing sucrose and ethylene glycol. However, for prepubertal testicular Results: 1. Sperm or spermatid were found totally in 35.3% (54/153) of non
tissues, it is considered necessary to re-evaluate the cryoprotectant and the obstructive azoospermic men using this optic system. 2. In 85% (54/64) of TESE
program of freezing, taking account of their being unlike adult testicular tissues. trials, spermatids and sperm were found when thick seminipherous tubules were
Moreover, testicular tissue from cancer patients may be contaminated with filled with homogenous fine and whitish granules that did not move so much
cancerous cells. It would be of importance to restore, with discretion in various within. 3. No spermatogenic cells (primary spermatocyte, spermatid) were found
respects, testicular cells collected from a pediatric cancer patient into that and sperms were absent (0/25) when seminipherous tubules were filled with
patient after completion of cancer treatment. lower density, yellowish, coarse granules. Yellowish substances were debris. 4.
Sperm or spermatogenic cells were likely to be found more frequently when the
308.1 – DOES A COMBINATION OF CHINESE HERBS AND ACUPUNCTURE whitish granules in the seminipherous tubules were not moving so much
TREATMENT AFFECT SPERM CHARACTERISTICS IN INFERTILE compared to those when they were moving more. 5. Pregnancy rates and
COUPLES? A PILOT STUDY miscarriage rates after microfertilization using testicular sperm and spermatid
were [38.9 % (7/18), 28.6% (2/7)], [22.2% (8/36), 37.5% (3/8)] respectively.
Ramon Velleman1,Tal Belo1, Ilya Barr, Guy Cassuto3, Shai Davids1, Yona Barak4.
1
Kibbutzim College of Education, Tel-Aviv, Israel; 2Israeli Fertility Center, Israel; Conclusions: With this system, it was possible to three-dimensionally visualize
3
Laboratory Drouot , France; 4Dr Yona Barak Laboratories LTD, Israel. the structure of seminiferous tubules and objectively evaluate the degree of
attachment of sperm cells, cellular density, cellular sizes, and color from the wall
Objective: Classic therapies are known to have a limited effect in the treatment thickness of the seminiferous tubules. Clinical results with this new photo
of patients with poor sperm characteristics. The aim of this study was to dynamic system will be improved.
determine the effect of the combination of Chinese herbs and acupuncture on
these males, who failed to conceive in their previous IVF-ICSI attempts
Design: Preliminary prospective analysis 308.3 – A NOVEL CRYOPRESERVATION TECHNIQUE FOR VERY FEW MOTILE
SPERM FROM SEVERELY INFERTILE MEN
Material and Methods: Sperm samples of 9 couples who failed to conceive in at
least 3 previous IVF-ICSI attempts, were analyzed by light microscope, according K. Kyono, H. Hattori, C. Nishinaka, M. Doshida, K.Yasuda, S. Kanto. Kyono ART
to WHO criteria. Herbs formulas were daily administrated. Acupuncture Clinic, Honcho, Aobaku, Sendai, Miyagi, Japan.
treatments, were done weekly. Sperm of severe oligozoospermia and azoospermia frequently demand freezing
Sperm analysis were performed before the treatment, 1 month and 2 months for future fecundity. However, no reliable protocol for freezing very low numbers
after the combined acupuncture treatments and herbs formulas started. A of sperm has been established. Therefore, we evaluated two new freezing
comparison of the following sperm parameters: volume of ejaculate, PH, sperm procedures for sampling very low numbers of sperm.
concentration, sperm motility and morphology, was performed before and during We used some ejaculated semen from infertile men. Sperm was retrieved by
the study. These parameters were also compared with sperm criteria of 12 putting a few sperm into a 2 μl droplet of cryoprotectant using a manipulator
patients, who underwent 2 sperm analyses within a 4 months period of time, under a reverse microscope. The droplet was diluted by mixing 0.7 of Sperm
without any conventional treatments, or any alternative treatment (control group). FreezeTM (FertiPro,N.V., Belgium)(SF) and 1.0 of HTF (Irvine, USA). Each included
Results: Out of the 9 couples, 3 conceived during 3 months time; two couples some sperm a 2μl droplet placed on 60 mm petri dish (Falcon1006, USA) was
conceived following IVF-ICSI treatment and the other following intrauterine covered with mineral oil and wrapped in Saran wrap. It was left for 10 minutes at
insemination. A tendency of 0.06 was noticed in a comparison of the rate of room temperature(RT), for 15 minutes at 4 , for 30 minutes on liquid nitrogen
normal forms before and after 1 month of treatments (14.7%±6.3 vs 28%±13.4, vapor, and immersed into LN2.
respectively). No change was noticed in the rate of normal forms in the control On the other hand, in the tip group, 2 μl droplets were aspirated into a CryotipR
group (28.67±12.3 vs 31.0±13.12%). (Kitasato Biopharma, Tokyo, Japan). It was left for 10 minutes at RT after closing
Conclusion: The combination of acupuncture and Chinese herbs may be a the tip point, for 5 minutes on LN2 vapor, and respectively and immediately
useful, no traumatic supporting treatment for males of couples which failed to immersed into LN2.
conceive in IVF, and intend to undergo further fertility treatments. Since our pilot In thawing procedure of the Dish group, the dish was thawed at RT until droplets
study is based on a small number of patients further investigation is still taking with sperm were completely thawed, and sperm was retrieved by a manipulator.
place. In thawing procedure of the Tip group, the cryotip was incubated in water of 37
for 20 seconds, and sperm was retrieved by a manipulator. We compared
308.2 – IMPROVEMENT OF A PHOTODYNAMIC SYSTEM FOR OBSERVATION OF retrieval rates of sperm (RR), survival rates (SR) and available useful rates (AUR)
SEMINIFEROUS TUBULES IN MICRODISSECTION-TESTICULAR SPERM for ICSI between the Dish group and the Tip group.
EXTRACTION (MD-TESE) In the Dish group, RR was 97.3% (299/309), and SR was 30.0% (82/273), and
Atsushi Tanaka, Motoi Nagayoshi, Shoichiro Awata, Norio Himeno, Izumi Tanaka. motility rate after thawing was 2.9% (8/275), and reaction of hypo osmotic
Dept. of Obstet. and Gynecol., Saint Mother Hospital, Fukuoka, Japan. swelling test (Host), positive was 27.9% (74/265), and AUR was 26.5% (82/309).
In comparison, in the Tip group, RR was 67.7% (239/353), and SR was 15.9%
Objective: Operating microscopes have long been used for microdissection- (36/226), and motility rate after thawing was 3.0% (7/232), and reaction of Host,
testicular sperm extraction (MD-TESE). Although “white, thick, and twisted” positive was 13.2% (29/219), and AUR was 10.2% (36/353). RR, SR, and AUR
seminiferous tubules have been targeted in TESE, it is necessary to observe the were significantly different between the two groups (P<0.01).
inside of the seminiferous tubules through the walls for more accurate
identification of sperm and sperm cells. From this point of view, we here In conclusion, it has been suggested that the freezing methods using a dish is a
investigated the potential of a newly modified photodynamic system. highly useful protocol for very low numbers of sperm. However, to raise the
collection rate and the survival rate we need to try other kinds of cryoprotectant
Methods: (1) Ophthalmology contact type lenses (2.0~3.0X, Blumenthal and improve the cryopreservation.
Suturelysis lens, VOLK OPTICAL INC. USA) were placed at the focus on the light
57 Geneva
Abnormal imprint establishment has been observed in the sperm of men

308.4 – ARGUMENTS TO IMPLEMENT THE SELECTION OF SPERMATOZOA AT with defective spermatogenesis. However, the exact mechanism underlying
HIGH MAGNIFICATION BEFORE ICSI this phenomenon remains elusive. The goals of the present study are 1) to
determine if the patient population in Montreal has imprinting defects in
H. Zech1, A. Stecher A1, M. Bach1 , T. Neyer1, M. Zinst1, P. Vanderzwalmen1,2, N. sperm DNA and 2) to elucidate the mechanisms underlying imprinting
Zech1. 1Institue for Reproductive Medicine and Endocrinology, Bregenz, Austria; defects in infertile men. We hypothesized that imprinting defects in human
2
Centre Hospitalier Inter Regional Cavell, Bruxelles – Braine l alleud, Belgium. sperm DNA may result from partial loss of function mutations in genes
Selection of spermatozoa during ICSI procedure is performed at a rather low involved in imprint establishment in the germ line.
magnification at 200-400X with Hoffman Modulation interferential contrast Materials and Methods: Patients with normal and defective spermatogenesis
(HMC) optics. Using such optical tools, selection of spermatozoa has severe were recruited in the McGill University Reproduction Centre. DNA
limitations. This has been one of the major concerns related to ICSI as the sperm methylation pattern of three imprinted regions (H19, SNRPN and IG
selection process, occurring during normal IVF in presence of cumulus cells, is differentially methylated regions (DMR)) were investigated in 15 patients.
totally bypassed. The changing of HMC by Nomarski differential interferential Six of these patients had male factor infertility and 9 had normal sperm
contrast and examination at high magnification, has allowed a better observation parameters. In parallel, to test our hypothesis, we tested the efficiency of
of sperm cells in real time. Using “motile-sperm organelle-morphology imprint establishment in the germ line of mice that carry heterozygous
examination” MSOME, “normal spermatozoa” exhibit a large panel of nucleus mutations in the genes known to be involved in imprint resetting and/or
malformations in terms of shape, size and presence of vacuoles are detected and spermatogenesis. We explored the effect of mutations in the 5,10-
normally not selected. However, a more acute selection of spermatozoa means methylenetetrahydrofolate reductase (Mthfr), DNA methyltransferase 1
that it is not always possible to find spermatozoa morphologically completely (Dnmt1), Dnmt3a, Dnmt3b and mutS homolog 5 (E. coli) (Msh5) genes. We
normal. Several studies reported that the existence of large vacuoles in the have also tested the effect of folate deficiency on DNA methylation in sperm.
nuclei of spermatozoa dramatically reduces the proportion of good quality
embryos reaching the blastocyst stage and logically the pregnancy (PR) and Results: incomplete methylation at the H19 DMR and partial methylation of
implantation (IR) rates. More over higher rates of abortion is also noticed. the SNRPN DMR were found in 3 and 4 patients, respectively. Consistent
with reports from other groups, the same defects were found in both males
As a consequence, the pending and crucial question concerns their meaning, with defective and normal spermatogenesis: among the patients with normal
origin and the further impact on embryo quality and downstream consequences spermatogenesis, two had partial methylation of the SNRPN DMR and one
on embryo development and more concerning, the progeny. had incomplete methylation of the H19 DMR. Among the patients with
Can we assume that there is a relationship between chromatin defect - DNA abnormal spermatogenesis, two had defects in H19 methylation and one had
damage and the presence of nuclear vacuoles? Two recent papers (Garolla 2008 partial methylation of the SNRPN DMR. Some of the mouse mutations were
and Franco 2008), reinforce the concept that an association between large associated with incomplete imprint establishment at the H19 locus and
vacuoles and secondary and tertiary DNA structure damages in sperm nucleus showed aberrant methylation patterns similar to those observed in the
exists. sperm of human males.
What are the consequences of DNA damage? In the light of the reports of Aitken Conclusions: Abnormal methylation at imprinted regions is not necessarily
(2007), the negative effect of sperm DNA fragmentation may affect the next associated with abnormal spermatogenesis in human males. Methylation
generation (Aitken, 2007). Furthermore a recent work of Fernandez-Gonzalez patterns reminiscent of those found in humans were observed for some of
(2008) in the mouse model demonstrated that DNA-fragmented spermatozoa in the mouse mutations, suggesting that partial deficiency for enzymes
ICSI can generate effects that emerge during later life, such as aberrant growth, involved in DNA methylation and/or meiosis may be the underlying
premature aging, abnormal behaviour, and mesenchymal tumors. mechanism of human imprinting defects.
Even though there is a strong “in vivo” selection after embryo transfer. we must This work was supported by funds from the McGill Reproductive Center
be cautious, in order to lower the potential risks mentioned here. (MRC) and the National Sciences and Engineering Research Council of
Canada (NSERC).
In fine, the one of the most frequent questions regarding IMSI relates to its
indications: should we perform IMSI to all the patients? Another debate raises the
question of what should we do for patients carrying 100% large vacuoles in their 309.2 – EFFECT OF MATURATION IN VITRO ON SPINDLE MORPHOLOGY IN
sperm samples. Even though there is no real proof in the human species on the HUMAN OOCYTES. A COMPARATIVE STUDY ON PATIENTS WITH MALE
abnormal outcome generated by spermatozoa carrying vacuoles, this ultra- FACTOR
morphology technique has to be added as an additional tool for ICSI knowing the
consequence of possible DNA damage for offspring (Carrell, 2008). Shall we M. Mette Munk, E. Novella-Maestre, F.B. Lindenberg, S.S. Jensen, S.
continue the attempt with his sperm or propose the option of sperm donor? The Lindenberg. Section for Reproductive Biology and ART, Nordica Fertility
establishment of new classification criteria, based on an assessment system, Clinic, Copenhagen, Denmark; Instituto Valenciano de Infertilidad (IVI),
seems a valuable approach to determine a threshold limit for making the right University of Valencia, Spain.
therapeutic decision. In conclusion, sperm selection before ICSI seems more and Introduction: Incomplete cytoplasmic maturation of in vitro matured (IVM)
more unavoidable whatever the screening technique (hyaluronique binding test), oocytes has been known to cause microtubule and filament alterations,
in order to lower the potential risks detailed here. Analysis of semen samples on which may results in abnormal pronuclear formation and later failed
an ultramorphologic scale for the presence of vacuoles should therefore be embryogenic development. In the present study we examined the influences
recommended to patients before ICSI. of in vitro maturation of human ova to the MFII stage in a standardized in
vitro maturation medium compared to in vivo matured MFII stage ova prior
to the ICSI procedure.
309.1 – SEARCH FOR MECHANISMS UNDERLYING IMPRINTING DEFECTS IN
SPERM Material & methods: A total of 44 patients admitted for IVM and 44 patients
admitted for IVF/ICSI were analyzed using a polscope for evaluating the
1,2,5 1 2 1
Anna K. Naumova , Sanny Moussette , Aabida Saferali , Nicola Dean , presence and placement of the spindle in relation to the polar bodies in all
Donovan Chan, Jacquetta Trasler2,3,4,5, Taiping Chen6, Teruko Taketo7, Rima MFII oocytes produced after egg collection prior to ICSI or after 28 h in vitro
Rozen2,3,5 , Seang Lin Tan1,5. Departments of 1Obstetrics and Gynecology, maturation following the IVM protocol.
2
Human Genetics, 3Pediatrics, 4Pharmacology and Therapeutics,
7
Department of Surgery and Biology, McGill University, Montreal, QC, Results: We found the same mean number of oocyte retrieved (IVM 4,9
Canada; 5Research Institute of the McGill University Health Centre, Montreal, oocyte versus ICSI 5,9 oocyte), the mean number of MFII oocytes after
QC, Canada; 6Novartis Institutes for BioMedical Research, Cambridge, MA, collection of IVF/ICSI oocytes were 5,1 versus 2,1 for the IVM oocytes after
USA. 28 h maturation (P<0,01) and finally we found a significantly higher rate of
displacement of the spindle apparatus in the in vivo matured oocytes
Introduction: Genomic imprinting is a parent-of-origin dependent epigenetic compared to the in vitro matured oocytes. (P > 0.05).
marking of chromosomes. During gametogenesis, genomic imprints are
erased, and reestablished depending upon the sex of the individual. Conclusions: These findings indicate that in vitro matured human oocytes
either mature from MF I to MFII normally having a normal spindle
April 19-22, 2009 58

appearance after 28 hours in vitro maturation or detoriate completely. This oocytes and granulosa cells. Hence, its expression in corpora lutea was
indicate, that IVM matured oocytes if they reach the MFII are normal, and localized on luteal cell membranes.
the obvious lower implantation rate in IVM might be due to other factors Conclusions: Our data demonstrate that PKC isotypes are differentially
such impaired endometrial development during the IVM procedure expressed in normal mouse ovary. The mode of action of DAG, Ca2+,
phospholipid dependent PKCα seems to be regulating the granulosa cell
activity and luteal cell functions. Oocyte maturation may also be maintained
via PKCa. However Ca2+ independent PKCδ and ε seems to regulate only
309.3 – PATERNAL AGE AFFECTS SPERM DNA FRAGMENTATION BUT NOT ITS
PACKAGING
luteal cell functions. Although we did not differentiate between the
Stéphanie Belloc, André Hazout, Martine Cohen-Bacrie, Moncef Benkhalifa, expressions of large and small luteal cells, localization of the novel PKC
Martine Dumont, Anne Marie Junca, Yves Menezo, Paul Cohen-Bacrie. isotypes PKCδ and ε support their role in luteolysis. We suggest that the
Laboratoire Eylau Unilabs, Paris, France. ovary took the advantage of differentially stimulated PKCs where they can
Introduction: Basic parameters such as concentration, motility and regulate different follicle stages and corpus luteum functions by processing
morphology are of limited value in determining the “embryotrophic” different PKC isotypes.
potential of sperm, i.e. the ability of sperm cell to establish a full term Acknowledgement: The study was supported by Akdeniz University,
pregnancy. DNA integrity, including secondary and tertiary structure i.e. Scientific Research Fund.
DNA fragmentation and decondensation are now mandatory to approach
this sperm functional aspect.
Material and Methods: Paternal age has only recently been considered 309.5 – IMPACT OF A GnRH ANTAGONIST ON THE OUTCOME OF CONTROLLED
(Klonoff-Cohen and Natarajan, 2004, Belloc et al. 2008), as affecting the OVARIAN HYPERSTIMULATION (COH) IN WOMEN WITH POLYCYSTIC
potential of sperm. We have tested on more than 1000 patients the effects of OVARY SYNDROME: A PROSPECTIVE AND RANDOMIZED STUDY
age on sperm DNA fragmentation (measured with TUNEL) and Laurel A. Stadtmauer1, Silvina Bocca1, Hind Baydoun2 , Beth Pultz1, Sergio
decondensaion measured with aniline blue (AB). AB stains lysine-rich Oehninger1. 1The Jones Institute for Reproductive Medicine, Department of
histone proteins, leaving the arginine-rich protamine proteins unstained and Obstetrics and Gynecology, Eastern Virginia Medical School; 2Department of
thus is more specific of sperm maturity. Epidemiology, Eastern Virginia Medical School; Norfolk, VA, USA.
Results: Sperm DNA fragmentation increases with age (p<0.001) but not Introduction: The study was conducted to evaluate the efficacy of the
with the percentage of atypical forms. In contrast, decondensation Gonadotropin releasing hormone (GnRH) antagonist Ganirelix® when used
decreases, but not significantly, with increased age (p=0.063), yet the quality as adjuvant to gonadotropin stimulation with recombinant follicle
of DNA packaging is negatively associated with the percentage of atypical stimulating hormone (Follistim®) in PCOS patients undergoing ovulation
forms (p<0.006). These two observations fit with the obvious less induction and IUI.
resistance towards Reactive Oxygen Species with age. ROS are involved in Materials and Methods: A prospective randomized controlled study of
DNA fragmentation but also in condensation through protamines cross anovulatory women with PCOS undergoing ovulation induction cycles with
linking. DNA damages affect the outcome of ART procedures. If the impact IUI (n=138). The 3 groups were: Group 1, Follistim® alone; Group
of fragmentation is evident, anomalies in the tertiary structure of sperm DNA 2,Follistim® with Ganirelix®, initiated at a follicle size ≥13 mm in diameter;
do affect, sooner or later, normal embryonic development even if fertilization Group 3,Follistim® with Ganirelix from day 1 of stimulation. The primary
is not impaired (Rousseaux et al. 2008). outcome measure was the clinical pregnancy rate per cycle of treatment.
Conclusion: These two parameters should not be neglected irrespective of Secondary outcomes measures included the total gonadotropin dose per
the age of the male partner, yet the age of the female partner can be cycle, number of days of treatment, peak serum estradiol levels in each
balancing aspect as the oocyte is able to repair partly fragmentation decay cycle, and premature luteinization rate and live birth rate per completed
and more weakly some tertiary structure anomalies (Ménézo et al. 2007). cycle..
Results: The results show a trend towards an improvement in the clinical
309.4 – THE LOCALIZATION OF SPECIFIC PROTEIN KINASE C ISOTYPES IN pregnancy rates per cycle initiated in patients treated with Ganirelix®
MOUSE OVARY (Group 2, 17/48, 35%), compared with Groups 1 (10/48, 22%) or Group 3
(8/42, 20%) with p=0.1. All except 2 pregnancies were singletons (2 twins in
Filiz Tepekoy, Zeliha Sahin, Gokhan Akkoyunlu. Department of Histology and Group 2). In the absence of Ganirelix® (Group 1), premature luteinization
Embryology, Medical Faculty, Akdeniz University, Antalya, Turkey. occurred more often (25% vs. 0% vs. 2.% p<0.05) and the LH levels on the
Introduction: The mammalian protein kinase C (PKC) consists of a family of day of hCG were significantly higher than in Groups 2 or 3. Group3 had the
twelve distinct members. They can be subdivided into three subfamilies highest cancellation rate and cost per cycle. Ganirelix® did not have any
which are classified on the basis of sequence similarities and their modes of effect on peak estradiol levels, total vials of r-FSH used or days of
activation. The conventional PKCs (α, β1, β2, γ) are all activated by stimulation.
phospholipids, in particular phosphatidyl serine, DAG (diacylsglycerol) and
Ca2+ where the novel PKCs (δ, ε, η, θ, µ) do not require Ca2+for its action.
Conclusions: Premature luteinization occurred commonly in women with
The atypical PKCs (λ, ι, ξ) respond neither to DAG nor to Ca2+ but they still
PCOS undergoing ovulation induction with Follistim ® alone and this is
associated with lower pregnancy rates. There may be a benefit in the
require phospholipids. The PKCs are suggested being critical in the addition of a GnRH antagonist. The higher, although not significant,
regulation of follicular development, ovulation and luteinization. Although it pregnancy rate in Group 2 patients suggests that the flexible Ganirelix
is hypothesized to control different cellular processes, the specific regimen could be the treatment of choice for PCOS undergoing
localization of these PKCs has not been enlightened in ovary up today. The gonadotropin stimulation and IUI.
results of this study will suggest the specific localization of PKC isotypes in
normal female mouse ovary.
Materials & Methods: Ovary samples were obtained from normal adult 309.6 – OVARIAN AGING: PROGNOSTIC FACTORS, BIOLOGICAL FACTORS AND
female mouse (n= 6). The localization of PKC isotypes was determined on OUTCOME OF ART
cryosections by using immunohistochemical techniques. Andrea R. Genazzani, P. Monteleone, O.M. Di Berardino, G. Simi, P.G. Artini.
Results: PKCα expression was present in all different maturation stage of University of Pisa, Pisa, Italy.
oocytes in developing follicules. The PKCα expression was also seen in Numerous evidences support the idea that the chief regulator of female
granulosa cells of Graaf follicules and steroidogenic luteal cells. reproductive aging is the ovary. Female ageing the most significant
The PKCδ expression was still apparent in luteal cells of corpora lutea. determinant of success in IVF. Ovarian functional decline with aging has
However, it was less obvious in the oocytes of Graaf follicules compared to been so far studied in depth in terms of accelerated depletion of the ovarian
PKCα expression and was negative in granulosa cells. follicle pool and reduced ability to produce oocytes competent for
fertilization and development. Few studies have addressed the question
The PKCε expression was showed a similar expression pattern with PKCδ in about potential causal factors of ovarian aging. According to the most
59 Geneva
relevant concept of ageing, age-associated malfunction results from transvaginally ultrasound guided follicle aspiration from a patient with
physiological accumulation of irreparable damage to biomolecules as an polycyctic ovaries (1,2).
unavoidable side effect of normal metabolism. Moreover, it was suggested Before the first successful birth with in vitro fertilization (IVF) and embryo
that an important environmental factor responsible for oocyte senescence transfer, the first culture and maturation of human oocytes in vitro were
might be represented by a reduced oxygen supply to the leading follicle, a carried out on oocytes that were obtained by laparatomy. Similar
condition dependent on a compromised perifollicular vascularisation. Future experiments were done by Steptoe and Edwards and they introduced
investigation of age-related molecular damage in the different ovarian laparoscopic method for aspirating oocytes from the Graff follicles (3).
components is imperative in order to develop strategies to possibly save or Laparoscopic oocyte retrieval procedure may be performed either 2 or 3
rescue the developmental potential of aged oocytes. puncture technique depending on the type of the laparoscope and the
experience of the surgeon. Each follicle is punctured and aspirated at an
309.7 – FIRST TIME EVIDENCE FOR INCREASED PLACENTAL GROWTH avascular site. The needle may be left in the follicle to permit flushing and
FACTOR MRNA IN ENDOMETRIUM OF PATIENTS WITH SUCCESSFUL reaspiration in case that an oocyte is not identified in the initial aspirate.
IMPLANTATION Following those first few births via laparoscopilly aspirated oocytes,
ultrasound guided oocyte collection techniques has been described. While
Alessandro Santi, Niklaus A. Bersinger, Dorothea Galié-Wunder, Micheal D. laparoscopic oocyte retrieval is generally performed under general
Mueller. University Hospital Berne, Berne, Switzerland. anesthesia with endotracheal intubation the major advantages for the TVS
Introduction: The aim of this prospective study was to analyse the in vivo technique include decreased exposure to general anesthesia, lower chance
vascularisation of the endometrium as described at hysteroscopy and to for operative complications, and feasibility of performing on an outpatient
determine a possible relationship with angiogenic factors and the basis. On the other hand laparoscopy identifies visible follicles on the
implantation rate. ovarian surface, whereas TVS also identifies intra-ovarian follicles.
Material and Methods: Consecutively admitted infertile patients with a There are many different ultrasound guided oocyte collection techniques.
planned hysteroscopic evaluation for infertility were asked to participate in The first follicular puncture under ultrasound guidance was described in
the study. The study protocol was approved by the local ethical committee. 1981 and the same group was introduced transabdominal transvesical
All patients had a pre-operative transvaginal sonography (TVS). To evaluate ultrasound directed oocyte recovery for the oocytes from the ovarian
the vascularisation of the endometrium at hysteroscopy the procedure was follicles a year later (4,5). Oocyte retrieval for IVF by ultrasonically guided
performed in the secretory phase of the menstrual cycle. The surgeon was needle aspiration via the urethra was proposed by Parsons and coworkers in
blinded to the TVS-findings. The quality of the endometrium was evaluated 1985 (6). The above techniques have in routine use during the days of
according to the Sakumoto-Masamoto grading (“good” vs. “poor”) at the conventional external transducers. Transvaginal sonography guided
time of hysteroscopy and an endometrium biopsy was taken at the end of transvaginal follicular aspiration was initially reported in 1983 by Gleicher et
the procedure using a soft curette. An aliquot of the biopsied tissue was al. and become dominant method over laparoscopy (7). In this technique
subjected to total RNA extraction, reverse transcription, and quantitative the follicles are punctured with a single or double lumen needle connected
polymerase chain reaction (QPCR) for several angiogenic factors. The data to the vaginal transducer with a guide. Aspiration with a negative pressure <
were analysed in order to find a possible relationship between the 20 Kpa (150 mm HG) is performed gently and there is no need to withdraw
vascularisation of the endometrium, the implantation rate (spontaneous the aspirating needle from the ovary to move the next follicle. This TVS
pregnancy, intrauterine insemination by husband and IVF/embryo transfer) guided follicular aspiration technique have become the method of choice
and the transcription rate (RT-QPCR) of the angiogenic factors. For during the years because of the refinements in instrumentation and
statistical evaluation a t-student test was applied using Instat Graph Pad increased experience of the clinicians. Within the years the automatic
version 4.0 for windows. aspiration and washing systems has been introduced and the follicles can be
aspirated and, if necessary, washed with various systems. It has been
Results: One hundred and sixty-two infertile patients with a median age of
shown that oocyte retrieval rates may be improved by using this flushing
36.4 years (range 26-43) were included in the study. One hundred and eight
systems in IVF cycles, especially in patients who have low number of
(66.7%) were classified endoscopically as having “good” mid-secretory
follicles (e.g. natural or minimal stimulation IVF cycles or in poor
endometrium and 54 patients (33.3%) as “poor”. There were no differences
responders).
in the distribution pattern of the infertility causes between these two groups,
the age of the patients, and the delay of infertility. The overall pregnancy rate Even transvaginal sonography (TVS) guided follicular aspiration has now
was 37.0% (60 patients). Endometrial biopsy samples from 16 patients with become the preferred procedure of choice for oocyte retrieval in IVM cycles
a successful pregnancy and a “good” endometrium (group A) were comparing with the conventional IVF oocyte retrieval it requires certain
compared with biopsies from 10 patients with a “bad” endometrium and modifications(8). In a typical IVM cycle oocyte retrieval was performed
without pregnancy (group B). The patients of group A presented a under ultrasound control between 10 and 14 days after withdrawal bleeding
significantly higher level of mRNA for Placenta Growth Factor (PLGF) mRNA using a specially designed aspiration needle and an aspiration pressure of
in the biopsy than those of group B (P = 0.0184), while VEGF-A, 7.5 kPa which is half of conventional aspiration pressure. Although
Angiogenins, and their different receptors did not show such any difference pregnancy rates have in general been relatively low, recent reports suggest
in normalised mRNA content between the two groups. improving IVM success by hCG priming (9). All the follicles had to be < 10
mm in diameter because data suggest that the presence of dominant follicle
Conclusions: This in vivo study demonstrates for the first time that PLGF in
at the time of immature oocyte retrieval is deleterious to outcome in IVM
the endometrium biopsy together with the hysteroscopic appearance
(10,11). The maximum diameter of the follicle on the day of oocyte recovery
(vascularisation) of the endometrium might be an important prognostic
provides one of the main differences between IVF and IVM cycles. A
factor for the evaluation of the success rate in a therapy for infertility. Our
specially designed 17-gauge single lumen aspiration needle was introduced
results need a larger number of patients in order to confirm this hypothesis.
into the follicle with the bevel facing downward to prevent loss of follicular
fluid. To prevent blood clotting heparinized saline was used as the aspiration
310.1 – EFFICIENCY OF IVM EGG COLLECTION medium. An IVM oocyte collection involves multiple ovarian punctures since
the single channel aspiration needle tends to block frequently when passing
Bülent Gülekli, Obstetrics & Gynecology and Division of Reproductive
through the dense ovarian stroma and follicular flushing is not performed.
Endocrinology & IVF Unit, Dokuz Eylul University School of Medicine, Izmir,
The collection also takes, on average, longer than IVF oocyte retrieval
Turkey.
because of the repeated flushing of the needle and the tubing in order to
In vitro maturation (IVM) of immature oocytes retrieved from women prevent the blockage of the needle (12,13). Because of the multiple ovarian
without any ovarian stimulation is a promising new treatment especially for punctures the IVM oocyte retrieval is likely to be more painful than a routine
women with polycystic ovary syndrome (PCOS), with many successful IVF oocyte collection. Consequently, the method of anesthesia used during
pregnancies reported worldwide. Although Cha et al reported the first birth an IVM oocyte collection, especially in some patients who have previous
using immature oocyte donation from oopherectomy specimens in 1991, in poor response with IV anesthetic, may become a general anesthesia.
1994 it was Trounson and colleagues who put IVM in the clinical practice
when they reported the first pregnancy using oocytes collected by
April 19-22, 2009 60

References
Cha KY, Koo JJ, Ko JJ, et al. Pregnancy after in vitro fertilisation of human 311.1 – PREIMPLANTATION GENETIC DIAGNOSIS (PGD) FOR CANCER
follicular oocytes collected from nonstimulated cycles, their culture in vitro PREDISPOSITION GENES
and their transfer in a donor oocyte program Fertil Steril 1991; 55: 109 Yuval Yaron, Tel Aviv, Israel.
Trounson A, Wood C and Kausche A. In vitro maturation and the fertilisation Inherited germ-line mutations in cancer predisposition genes are
and developmental competence of oocytes recovered from untreated responsible for about 10% of cancers and may explain aggregation of
polycystic ovarian patients Fertil Steril 1994: 62; 353 cancer cases in some families. Such mutation may involve oncogenes,
Steptoe PC and Edwards RG. Birth after reimplantation of human embryo. tumor-suppressor genes, mismatch-repair genes, and others. Once a
Lancet 1978; 2: 366 pathogenic mutation has been identified, it is possible to perform pre-
Lenz S, Lauritsen G, Kejlow M. Collection of oocytes for IVF by ultrasonically symptomatic testing, as well as offer prenatal diagnosis and termination of
guided follicular puncture Lancet 1981; 1: 1163 affected pregnancies. This however poses significant psychological, ethical,
moral and legal issues. Alternatively, it is possible to perform
Lenz S, Lauritsen JG. Utrasonically guided percutaneous aspiration of preimplantation genetic diagnosis (PGD) in single cells from embryos
human follicles under local anesthesia: A new method of collecting oocytes obtained by in vitro fertilization (IVF). Transfer of embryos free of the
for in vitro fertilization. Fertil Sterl 1982; 36:673 mutation will ensure birth of unaffected offspring. While circumventing
Parsons J Riddle A, Booker M. Oocyte retrieval for in vitro fertilization by some of these issues, PGD for cancer predisposition raises new dilemmas,
ultrasonically guided needle aspiration via uretra Lancet 1985; i: 1076 and according to some opinions, presents the "slippery slope" towards
"designer babies". In this talk I will discuss these issues vis-à-vis our clinical
Gleicher N, Friberg N, Fullan N et al. Egg retrival for in vitro fertilization by
experience.
sonographically controlled vaginal culdocentesis Lancet 1983; 2: 508
Gülekli B,Demirtas E, Buckett WB. Immature oocyte collection. In In-vitro
Maturation of Human Occytes Basic sciences to clinical application Edited by 311.2 – PGD IMPROVED PGD TESTS AND CLINICAL APPLICATIONS
Tan SL, Chian R, Buckett WB,pp 253-263 , 2007 InformaUK Ltd René Frydman, Service de Gynécologie-Obstétrique et Médecine de la
Chian RC, Gülekli B, Buckett WM, Tan SL. Priming with human chorionic Reproduction, Hôpital Antoine Béclère; Univ Paris-Sud ; INSERM U782;
gonadotropin before retrieval of immature oocytes in women with infertility Clamart, France.
due to the polycystic ovary syndrome N Engl J Med 1999; 341: 1624 Preimplantation genetic diagnosis (PGD) is used to analyze embryos genetically
Russell JB. Immature oocyte retrieval combined with in-vitro oocyte before their transfer into the uterus. It was developed first in England in 1990, as
maturation. Human Reprod. 1998;13:63 part of progress in reproductive medicine, genetic and molecular biology. PGD
offers couples at risk the chance to have an unaffected child, without facing
Cobo AC, Requena A Neuspiller F et al. Maturation in vitro human oocytes
termination of pregnancy. Embryos are obtained by in vitro fertilization with
from unstimulated cycles: selection of the optimal day for ovum retrieval
intracytoplasmic sperm injection (ICSI), and are biopsied mostly on day 3;
based on follicular size. Hum Reprod 1999:14;1864
blastocyst biopsy is mentioned as a possible alternative. The genetic analysis is
Child TJ., Abdül-Jalil AK., Gülekli B., Tan SL., In vitro maturation and performed, on one or two blastomeres, by Fluorescent In Situ Hybridization (FISH)
fertilisation of oocytes from unstimulated normal ovaries, polycystic ovaries for cytogenetic diagnosis, or Polymerase Chain Reaction (PCR) for molecular
and women with polycystic ovary syndrome. Fertil Steril 2001; 76:936 diagnosis. Genetic analysis of the first or second polar body can be used to study
Child TJ, Phillips SJ, Abdul-Jalil AK, Gülekli B, Tan SL. A comparison of in maternal genetic contribution. Only unaffected embryos are transferred into the
vitro maturation and in vitro fertilization for women with polycystic ovaries uterus. To improve the accuracy of the diagnosis, new technologies are emerging,
Obstet Gynecol 2002; 100: 665 with Comparative Genomic Hybridization (CGH) and microarrays.
In Europe, depending on national regulations, PGD is either prohibited, or allowed,
or practiced in the absence of recommendations. The indications are
310.3 – HCG PRIMING FOR IVM
chromosomal abnormalities, X-linked diseases or single gene disorders. The
Weon-Young Son, Royal Victoria Hospital, Montreal, Canada. number of disorders being tested increases. In Europe, data collection from the
A major side-effect of controlled ovarian hyperstimulation (COH) in patients year 2004 reports that globally 69.6% of cycles lead to embryo transfer and
with polycystic ovary or polycystic ovarian syndrome (PCOS) is the risk of implantation rate is 17%. European results from the year 2004 show a clinical
ovarian hyperstimulation syndrome (OHSS). In-vitro maturation (IVM) of pregnancy rate of 18% per oocyte retrieval and 25% per embryo transfer, leading
immature oocytes represents a potential alternative for the fertility treatment to 528 babies born. The cohort studies concerning the paediatric follow-up of
of these patients. Recently, applications of FSH (hMG) or hCG priming have PGD babies show developmental outcomes similar to children conceived after
been used before immature oocyte retrieval to improve the success rate of IVF-ICSI.
IVM procedures. Recent advances include Human Leucocyte Antigen (HLA) typing for PGD
Although it is still controversial, hCG priming before oocyte retrieval seems embryos, when an elder sibling is affected with a genetic disorder and need stem
beneficial in terms of easier oocyte retrieval, easier oocyte identification cell transplantation. The HLA-matched offspring resulting can give cord blood at
under stereomicroscope, maturation competence, and may increase the birth. Preimplantation genetic screening (PGS) consists in euploïd embryo
harvest in vivo mature oocytes. Recently, new approaches attempted to selection; it could be used for advanced maternal age, repeated implantation
improve IVM program such as extending hCG stimulation or optimal hCG failure, single embryo transfer or idiopathic recurrent pregnancy loss. These
timing before immature oocytes retrieval. Therefore, as a first option, hCG- applications are controversial. PGD for inherited cancer predispositions is
priming IVM treatment can be offered to women with PCO(S) instead of discussed and social sexing remains prohibited in Europe.
conventional IVF treatment with ovarian stimulation. In this session, I would Conclusion: PGD requires a close collaboration between obstetricians, fertility
like you to introduce on embryological factors affecting the clinical outcome specialists, IVF laboratory and human geneticists. It needs intensive effort,
of IVM cycles after hCG priming. expensive techniques and is demanding for the patients, but it offers tremendous
opportunity for couples whose previous child has exhibited genetic abnormalities.
The debate on certain indications is ongoing.
61 Geneva
Prophase I (PI) germinal vesicle present, 33 metaphase I (MI) no first polar

312.1 – ULTRASOUND GUIDED VERSUS BLIND TOUCH EMBRYO TRANSFER: body and no germinal vesicle, 10 metaphase II (MII) first polar body
THE EGYPTIAN EXPERIENCE present. Immature oocyte (PI) and (MI) were cultured to capable of
undergoing final in vitro maturation. 15 primordial follicles isolated from
O. Azmy, T. Taha, M Bibars, M. Refaat. Reproductive Medicine Department, ovarian tissue were also analysed.
National Research Centre, Cairo, Egypt.
For each group the minimum of the transmission (arbitrary unit, a.u.) and
Objective: To compare the success rate of ultrasound-guided (USG) embryo corresponding wavelength position (nm) of the spectra were computerized
transfers versus clinical touch (CT) embryo transfers among patients and compared with T test of Student and ANOVA.
undergoing ICSI cycles.
Results: The 4 oocyte groups presented different transmission spectrum
Design: A case-control study profiles. Prophase I, Metaphase I and Metaphase II have significant
Setting: analysis of women records undergoing fresh embryo transfer at the minimum of transmission (0.34, 0.44, 0.49 a.u. respectively - p<0.05) and
National Research Centre in the Department of Reproductive Medicine, over are characterized by a progression of their transmission spectrum with the
three year period between 2004 and 2007. degree of maturity. Primordial follicles presented with a different white light
transmission profile (0.21 a.u.).
Patients: Eight hundred and fifty three cycles of ICSI were analyzed as
regards the technique of embryo transfer, whether by USG or the CT Conclusions: Transmission spectrum analysis of oocyte by using of this
technique. microsystem, a non deleterious technology allows the assessment of
cytoplasm maturity degree during ICSI attempts. This technology could be
Methodology: Comparisons were made as regards the chemical and clinical
an alternative to microscope observation. Moreover, it could play a role
pregnancy rates as well as the ectopic pregnancy and miscarriage rates
during follicle culture or oocyte in vitro maturation protocols.
between the two groups. Corrections were made for the patient’s age, cause
of infertility, body weight, induction protocol, number and quality of
embryos transferred and the difficulty or blood stained procedure. 312.3 – HYSTEROSCOPIC ENDOMETRIAL EMBRYO DELIVERY (HEED):
Results: The chemical pregnancy was significantly higher among the USG TRANSFER OF DAY 2-5 EMBRYOS
versus the CT embryo transfer techniques (47.6%, 25.5% respectively, Michael Kamrava1, Asha Bhargava2, Jerry Hall3; 1West Coast IVF Clinic,
P<0.001). Furthermore, the clinical pregnancy was higher among the group Beverly Hills; 2LA Center for Embryo Implantation, Beverly Hills; 3UCLA
using the transvaginal USG during embryo transfer compared with those in Geffen Medical School, Los Angeles; CA, USA.
the group using the clinical CT method (32.3% and 12.3% respectively,
P<0.0001). In-spite of the increased inconvenience a woman may Introduction: Since the inception of in vitro fertilization (IVF), the procedure
experience during the abdominal ultrasound with embryo transfer, however, has seen many advances that have significantly improved pregnancy rates
the woman has more than double the chances of being pregnant if the as well as a reduction in complication rates. Embryo transfer at the cleavage
embryos were transferred under vision rather than blindly (Relative risk stage began as the standard of practice both because of limitations in the
1.86; 95%CI 1.53 to 2.25 , P<0.0001). Nevertheless, there were no ability to culture later stage embryos at that time and because of the ability
significant differences in ectopic pregnancy or miscarriage rates between of human embryos at that time to develop even at an early stage when
the two groups. placed into the uterus. However, improved understanding of embryo
metabolism led to the development of a culture sequence that optimizes
Conclusion: In our experience, although patient’s discomfort is more while growth and implantation at various stages of development, including
using the USG technique ET, we believe that this maneuver significantly blastocysts.
improves the chances of achieving a successful pregnancy.
The benefits of blastocyst stage transfer have been established for the
routine blind transfer technique of catheter introduction into the uterine
312.2 – A MICROSYSTEM WITH WHITE LIGHT OPTICAL SENSOR TO QUALIFY cavity when a patient produces a large number of oocytes or has proven
THE CYTOPLASM MATURITY OF HUMAN OOCYTES capacity to produce blastocysts. Higher pregnancy and live birth rates are
observed for other than cleavage stage embryos, especially in patients with
Florie Vidberg1, Clotilde Amiot1, Christian Pieralli 2, Bruno Wacogne2, Alphée
high numbers of eight-cell embryos present 72 hours after fertilization. This
BAILLY1, Christiane Joanne1, Germain Agnani3, Christophe Roux1. 1Service
also implies improved success with the transfer of fewer embryos, which
de Génétique Histologie Biologie du Développement et de la Reproduction
may translate into fewer multiple gestations and the ability to implant the
CHU; 2Institut FEMTO-ST; 3Service de Gynécologie Obstétrique CHU;
most viable embryo(s) available. However, recent recommendations in using
Besançon, France.
lower medication dosages for the controlled ovarian hyperstimulation, and
Introduction: During ICSI attempts oocytes reaching metaphase II are in patients with lower response or advanced age, the number of developing
microinjected. A morphological examination under a microscope provides embryos are limited and cleavage stage embryo transfers may be more
the usual means for determining oocyte maturity based on the meiotic advantageous. Also, since a significant number of patients embryos develop
status. On the other hand, cytoplasmic maturity is difficult to assess using poorly beyond day three, early transfer is clinically prudent.
this method. In this study the cytoplasm maturity was analysed using a
In this study, we use a flexible mini-hysteroscope with a flexible catheter for
microsystem recording of white light transmission spectra of oocytes.
direct delivery of embryos onto the endometrium under direct visualization.
Materials & Methods: The microsystem consists of an optical sensor We have previously shown that our technique for blastocyst stage
integrated onto a silicon anodic bonded lab-on-chip (LOC). This LOC was implantation resulting in a high pregnancy outcome. Now we seek to explore
initially devoted to micro fluidic application for moving and trapping a single the outcomes of embryo transfer at the cleavage stage versus the blastocyst
oocyte. In the present study the oocyte is held with a micropipette. stage, using our novel approach. We hope to further optimize pregnancy
Two aligned optical fibres were connected to the LOC surface. A white light and birth rate outcomes and thereby minimize the risk of multiple births and
source is launched into the illumination fibre (50 µm core diameter). The complications.
light propagates through the oocyte and is collected with another fibre (100 Objectives: To compare pregnancy and birth outcomes of embryo transfers
µm core diameter). The transmission spectrum of the oocyte is recorded performed on day two or three or five performed under direct visualization.
and divided by the transmission spectrum of the IVF medium only.
Materials and Methods: 32 consecutive patients with Infertility of various
The reference transmission spectrum I1(λ) is given by I1(λ) = I0(λ)C1R origins underwent Hysteroscopic Endometrial Embryo Delivery (HEED) on
where I0(λ) is the illumination spectrum, C1 the coupling coefficient day two or three or day five after fertilization. Controlled ovarian
between fibres and R(λ) the spectra response of spectrophotometer. The hyperstimulation was done using standard protocols. Transvaginal oocyte
collected spectrum is given by I2(λ)=I0( )T(λ)C2R(λ) with T(λ) oocyte retrieval was performed under local anesthesia with mild sedation. All
transmission spectrum and C2 the new coupling coefficient between fibres women received some type of luteal support, be it progesterone or hCG.
due to light focalisation by oocyte. The global spectrum is: Oocytes were fertilized and cultured in early cleavage medium (Irvine
I2(λ)/I1(λ)=(C2/C1)T(λ). Scientific) at 37 degrees C and 5% CO2 in air. Embryos were transferred at
Oocytes excluded from ICSI attempts were tested after denudation: 57 48-120 hours post fertilization.
April 19-22, 2009 62

Results: Table 1. Pregnancy Outcomes by Day of Transfer

Day 2 Day 3 Day 5 312.4 – NOVELHAND FREE ULTRASOUND GUIDED EMBRYO TRANSFER
Average Age (Years) 39 35 37
METHOD USING NEW STABILIZING DEVICE
Total number of patients 16 13 3

A. Rahim Hallob, Basildon University Hospital and Brentwood Fertility
Centre, Essex, UK.
Number of cancellations 0 0 0
Embryo Transfer is a major and most important step of Invitro Fertilisation
Number of patients with retrieval 16 13 3 and Intracytoplasmic Sperm Insemination Treatment Cycles. The new
% Total pregnancies per retrieval 40.0 61.5 66.7 Method will shorten the time needed for ultrasound guided ET, prevent
unnecessary manipulation,and free the operator to carryout other needed
% Spontaneous abortions of total pregnancies 0 25.0 0
tasks during ET such as second person check. The new stabilizing and fixing
% Multiple pregnancies of total pregnancies 16.7 0 50.0 device allows the outer catheter to be fixed in place on to the speculum,
% Ectopic pregnancies 0 0 0 therefore, minimising intrauterine movements of the tip of the outer and
inner catheters during ET and reduce truama to the endometrium. Its
% Ongoing live births per transfer 33.3 30.8 66.7
benefits come at a relatively cheap cost, as well as being user friendly.
Our results show that the percent of total pregnancies per retrieval was
highest on day of transfer five, though this is not significantly higher than
the percent on day of transfer three or two due to the limited number of
patients transferred on day five. The percent of total ongoing live births per 312.5 – CERVICAL MUCUS STATUS CAN BE ACCURATELY ESTIMATED BY
transfer is not different between transfer on day two and three, and percent TRANSVAGINAL ULTRASONOGRAPHY DURING FERTILITY EVALUATION
of ongoing pregnancies occurring with transfers on day five is higher than
Igal Wolman, Tamar Birenbaum Gal, Ariel J Jaffa. Ultrasound Unit in
transfers on day three or two. (Chi square value for day two vs. five = 1.36,
Obstetrics and Gynecology Lis Maternity Hospital, Tel-Aviv Medical Center,
0.5<P<0.1; chi square value for day three vs. five = 1.34, 0.5<P<0.1).
Tel-Aviv, Israel.
Of pregnancies that occurred with transfer on day three, fifty percent
Objective: To correlate the diameter of the cervical canal determined by
resulted in miscarriage or biochemical pregnancy, while only sixteen percent
transvaginal ultrasonography (TVS) to clinical assessment of the cervical
had this result when transfer occurred on day two. Only one multiple
mucus.
pregnancy occurred with day of transfer two and five, while none of the
transfers on day three produced a multiple pregnancy. Finally, no ectopic Design and Setting: Prospective study in an academic ultrasound unit.
pregnancies were observed in either group. Patients: Women (n = 101) undergoing an infertility workup or treatment.
Conclusions: These results using Hysteroscopic Endometrial Embry Delivery Interventions: Ovulatory cycle evluation.
(HEED) show for the first time that the technique is very effective with both
cleavage stage embryos and blastocysts, including day two embryos. These Main Outcome Measure(s): Cervical diameter measurements made from
results are even more remarkable due to our clinical bias to replace embryos TVS images were correlated to a cervical mucus score which was assessed
on day 2 when presented with poor quality or low numbers of embryos. Our by a different observer.
findings could offer new hope of success for a large number of poor Results: The cervical canal diameter values correlated well with the
prognosis IVF patients. calculated cervical mucus scores. The mean ± SE cervical canal diameter
References: was 0.9 ± 0.1 mm for women with a low (≤5) and 2.1 ± 0.1 mm for women
with a high (>5) cervical mucus score (P<0.001). A cervical canal diameter
Blake DA, Farquhar CM, Johnson N, Proctor M. Cleavage stage versus of 1 mm emerged as a single predictor of favorable versus hostile cervical
blastocyst stage embryo transfer in assisted conception. Cochrane Database mucus, with a sensitivity and specificity of 83.7% and 80.8%, respectively.
of Systematic Reviews 2007, Issue 4. Art. No.: CD002118. DOI: 10.1002/
14651858.CD002118.pub3. Conclusion(s): TVS, which is routinely used as a tool for follicular growth
monitoring in patients undergoing ovulation induction, might also
Practice Committee of the American Society for Reproductive Medicine. simultaneously be used for estimating cervical mucus measurements. With
Blastocyst production and transfer in clinical assisted reproduction. Fertil no additional effort or expense, these data may help to optimize individual
Steril. 2004 Sep;82 Suppl 1:S149-50. patient management.
Schoolcraft WB, Gardner DK. Blastocyst versus day 2 or 3 transfer. Seminars
in Reproductive Medicine 2001; 19:259-268.
63 Geneva
312.6– ULTRASOUND MONITORING AND HORMONAL PROFILES IN SERUM 312.7 – A STUDY OF EPIGENETIC ALTERATIONS ASSOCIATED WITH
AND FOLLICULAR FLUID FOR UNSTIMULATED IVF-CYCLES IN PREMATURE OVARIAN FAILURE
RELATION TO “EMPTY FOLLICLE SYNDROME” Manda Ghahremani1, Courtney Hanna2, Luana Avila2, Maria Penaherrera2,
Mikael Tang-Pedersen, Lars Westergaard. Fertility Clinic, Odense University Karla L Bretherick2, Margo R Fluker1-2, Wendy P Robinson2. 1Department of
Hospital, Faculty of Health - Institute of Clinical Research, University of Southern Obstetrics & Gynecology, 2Department of Medical Genetics, University of
Denmark, Odense, Denmark. British Columbia, 3Genesis Fertility Centre, Vancouver, BC, Canada.
Introduction: Over the years one observation remains constant in Introduction: Premature ovarian failure (POF) affects 1% of women with a
unstimulated / natural cycle IVF. That is the approx. 20 % of so called largely idiopathic and poorly understood etiology. The objective of this study
“empty follicles”. Empty follicle syndrome (EFS) has been debated, but since was to identify specific epigenetic alterations by measuring DNA methylation
it seems a constant finding a deeper understanding of the follicular of gene regulatory regions in women with POF vs. controls.
maturation process is desirable. In this study we investigate hormonal levels Materials and Methods: Blood samples were collected from idiopathic POF
in follicular fluid (FF) and serum for unstimulated IVF-treatment of ± patients (amenorrhea for at least 3 months and 2 serum FSH levels of >40
oestradiol-primed patients from start of the cycle till day of oocyte pick-up mIU/ml obtained > 1 month apart prior to age 40) and control women (CW)
(OPU), along with ultrasonic measurements of follicle size and number and (healthy pregnancy after age 37 with out a pregnancy loss). Genomic DNA
endometrial thickness. was extracted from EDTA anticoagulated blood and bisulfite converted for
The aim is to identify possible correlations between endocrinology, analysis using the Illumina Golden Gate Methylation Panel which measures
ultrasound monitoring and failed cycles due to “EFS”. DNA methylation at 1506 CpG sites in the promoter regions of 807 genes in
Materials And Methods: This retrospective analysis includes 68 women 18 POF and 21 CW. Candidate genes with altered epigenetic marks between
under the age of 37, referred to IVF-treatment due to tubal, male or POF and CW were identified based on 3 selection criteria: a nominal p-value
unexplained factor, and with regular cycles from 26-34 days and FSH <15 <0.05, absolute methylation difference of >5% and False Discovery Rate
U/L. Ultrasound (US) examination and blood samples were undertaken (FDR) of <50% using the “Significance of Analysis of Microarrays” (SAM)
during the early, mid and late follicular phases until the appearance of a version 3.01. Genes of interest were further analyzed for quantitative
dominant follicle > 16 mm, with a corresponding endometrium > 8 mm, methylation at specific CpG sites using pyrosequencing in 30 POF and 30
when an hCG-injection of 6500 IU was administered 34 hours prior to OPU. CW.
The aspiration of the mature follicle provides FF for measurements oflevels Results: After comparison of DNA methylation profiles of POF and CW using
of AMH, progesterone, oestradiol, androstendione and testosterone. The the above mentioned criteria, 16 genes met our criteria as candidate genes
mature oocyte is fertilized and transferred 2 days post-OPU. with altered epigenetic marks in the POF group. All of these genes were
Results: A total of 68 patients underwent 70 cycles of unstimulated IVF. OPU hypermethylated in the POF group as compared to CW. In addition, the
was cancelled in13 of these due to premature ovulation. Of the remaining 55 Androgen Receptor (AR) gene, which also was hypermethylated, met the
patients who went through OPU, 12 had an “empty” large follicle and from last two selection criteria and was followed up due to its functional role in
45 women a mature oocyte was retrieved. Of these 30 were fertilized and 26 the ovary. To further validate these results, DNA methylation of the AR gene
transferred at 4-cell stage. FF-concentration of progesterone in the12 cycles was quantified by pryosequencing in a larger group of POF and CW.
with EFS were significantly lower than in the 45 cycles with retrieval of a Pyrosequencing showed significantly higher DNA methylation of the AR
mature oocyte (9.5 ± 2.2 vs. 13.5 ± 1.1µg/ml; p = 0,042), despite the fact promoter region in POF vs. CW (p=0.007).
that US- criteria were met. Ratio of E2/P4 was significantly higher in EFS. Conclusions: This is a novel human study identifying epigenetic alterations
Compared to OPU cycles with retrieval of a mature oocyte, serum-LH levels in POF. 16 genes were found to be hypermethylated in POF compared to CW
on day of OPU was significantly lower for EFS ( p=0,0413) whereas se-FSH and several of them have a functional relevance in follicular development or
and se-E2 were similar in the two groups. Ratio s-(E2/LH) is also autoimmune processes. The further validate and quantify methylation of
significantly lower (p=0,0363) confirming a trend towards a slightly higher these genes, pyrosequencing experiments will be carried out, as was done
s-E2 in EFS. for the AR. The hypermethylation of the AR gene in POF patients may cause
Follicular mean diameter and endometrial thickness on last day of decreased level of the AR in these women. This is especially interesting
monitoring were similar in EFS and cycles with successful OPU. given a recent report of induced POF in AR deficient mice. Specific
epigenetic markers, as identified by DNA methylation array profiling in
Conclusions: Cycles with EFS exhibit different hormonal profiles than cycles blood, may serve as useful biomarkers for POF and other fertility disorders.
with mature oocytes retrieved. This indicates some degree of immaturity However, it will need to be determined if these methylation changes are
and/or a less capacitated oocyte-cumulus complex. However this is not present prior to diagnosis, or are a consequence of menopause itself.
visualized by standard 2-D ultrasound monitoring.
April 19-22, 2009 64

ORAL ABSTRACTS – WEDNESDAY
from day 3 of the period to the day when the maturation trigger is
400.1 – DERIVATION AND CHARACTERIZATION OF HUMAN EMBRYONIC STEM administered. Instead of an hCG injection, we utilize Gn-RhH agonist spray
CELLS as a maturation trigger to induce the LH flare up. Then, we aspirate oocytes
33 hours later.
Outi Hovatta, Karolinska Institutet, Karolinska University Hospital Huddinge,
Sweden and Geneve University Hospital, Switzerland. Single embryo transfer based on the Intra-tubal Embryo Recurrence Theory
Since the first outgrowths of cells from the inner cell mass of human Most importantly, we have been carrying out single embryo transfer for all
embryos (Fishel et al. Science 1986, Bongso et al. Hum Reprod 1998), and IVF cycles for the last 5 years. As a result, our multiple pregnancy rate has
the first permanent hESC lines (Thomson et al. Science 1998, Reubinoff et dropped to around 1 %. Along with this, the incidence of ectopic
al, Nature Biotechnol 2000), an intensive research period of derivation, pregnancies has decreased in our clinic, due to the fact that we provide
characterization, culture and differentiation of these cells has evolved. different treatment for women with tubal infertility to that we give those
Because of the potential of hESC in cell therapy, differentiation protocols to without the tubal infertility factor. Specifically, we treat women with tubal
many cell types, such as several cell types of nervous tissue, infertility with blastocyst transfer, and women without the tubal factor with
cardiomyocytes, hepatocytes, hematopoietic stem cells, bone, and many day 2 ET. Based on this result, we believe the zygote which has been
other cell types have been developed. It has been difficult in human to returned to the uterus moves to the Fallopian tube and develops in the tube
obtain clean populations of certain single cell types. Thinking of cell up to the blastocysts, then returns back to the uterus where it implants
transplantation this is a problem since non-differentiated cells among the itself. This has been shown in our animal studies. These animal studies,
populations are tumorigenic. Even a single pluripotent cell can give rise to a along with the human experience, suggest that blastocyst transfer may be a
teratoma. Before clinical trials this problem needs to be solved. Comparative solution for implantation failures in women with tubal factor infertility.
genomic hybridization and single nucleotide polymorphism profiling have
shown potentially malignant changes in these cells in culture. 402.2 – EMBRYONIC STEM CELLS DERIVED FROM PGD-AFFECTED EMBRYOS
Establishing clinical grade hESC has been another goal. There are strict FOR MODELING HUMAN GENETIC DISORDERS
requirements for products which will be accepted for clinical use. Good Dalit Ben-Yosef, Racine IVF Lab, Scientific and Managerial Director, Lis
manufacturing practice (GMP) quality system is required by both European Maternity Hospital, Tel-Aviv Sourasky Medical Center, Tel Aviv University, Tel
Medicines Agency (EMEA) and Food and Drug Administration (FDA). Aviv, Israel.
Optimal clinical grade cells have not been in contact with any animal derived
substances, because they are immunogenic and may contain microbes. The Human embryonic stem cells (HESC) carrying specific mutations can be
lines have to be derived and cultured from the beginning according to good used as a valuable tool for studying genetic disorders in human. One
to the quality system. Our recent defined feeder cell free culture system favorable approach to obtain such mutant HESC lines is their derivation
using recombinant human laminin 511 (Rodin et al. 2009) and chemically from affected preimplantation genetic diagnosed (PGD) embryos. These
defined culture medium has facilitated achieving this goal. The unique cells are especially important for modeling human genetic disorder
immunogenicity of hESC is another question which has to be solved. for which no good research models currently exist. They can be further used
to gain new insights on developmentally regulated events that occur during
Establishment of human induced pluripotent cells (Takahasi and Yamanaka human embryo development and that are responsible for the manifestation
2007), further development of such cells questioned the need of hESC any of genetically inherited disorders. In addition they have a great value for the
more. However, too little is known about these cells to say that really can be exploration of new therapeutic protocols, including gene therapy-based
used in human cell replacement. Possible genetic and epigenetic problems treatments and disease-oriented drug screening and discovery.
caused by the reprogramming are to be solved, even though non-viral
transduction will be likely to be achieved. All the knowledge accumulated The lecture will focuse on the importance of deriving HESC lines from
regarding the characterization of hESC and their differentiation is already genetically abnormal embryos. It will describe the first report on using PGD-
now applicable to human iPS cells, even though there appears to be also derived HESC for modeling a developmentally regulated genetic disease –
differences. For the time being, both cell types need to be studied. Fragile X.
401.3 – MINIMUM STIMULATION 403.1 – LAPAROSCOPIC TREATMENT OF UTERUS BICORNIS
Osamu Kato, Kato Ladies Clinic, Tokyo, Japan. Bruno J. van Herendael, Endoscopic Training Centre Antwerp (ETCA),
Gyntech BVBA, Antwerp, Belgium.
Shift from conventional IVF to minimal stimulation IVF
The topic concerns the laparoscopic technique of the classical Strassmann
Our clinic has become the largest IVF center in the world. In 2008, the operation for bicornate uteri. The second look hysteroscopy and
number of the IVF cycles carried out by us reached 20,000. For the last 14 laparoscopy prove that the technique is feasible and yields good results with
years, we have gradually shifted our method of IVF from conventional IVF to a minimal discomfort for the patients.
minimal stimulation IVF. Recently, we have also been performing natural IVF
cycles, and the results are very encouraging.
In the past, like most IVF centers in the world, we mainly performed 403.3 – ROLE OF HYSTEROSCOPY IN INFERTILITY AND IVF
conventional IVF using large dose of gonadotropin. This caused many side Luca Mencaglia, Geneva University Hospitals, Florence, Italy.
effects and complications such as Ovarian Hyperstimulation Syndrome
In the “”office hysteroscopy” the introduction of mini-endoscopes makes
(OHSS) and multiple gestations. These have been and still are both medical
hysteroscopy even more simple and painless. The technique is an office
and social problems.
procedure. We consider the “office hysteroscopy” a well tolerated innocuous
For the last 10 years, we have mainly used the Clomiphene, with or without procedure which could be considered as a routine procedure in all the cases
HMG protocol, which was developed in our center. This has significantly of infertility. A high level of expertise is not a prerequisite to performing
reduced the side effects of medication, and also reduced the patients’ hysteroscopy on an outpatient basis.
financial burden.
Hysteroscopy is able to detect important unsuspected endouterine
Our standard protocol for the minimal stimulation IVF using Gn-RH agonist abnormalities in 18% of patient with two failed IVF-ET and in whom HSG did
as an oocyte maturation trigger not show any abnormality.
In the controlled ovarian hyper-stimulation method, which is now popular all Indication for “office hysteroscopy” in ART
over the world, intra-muscular injection of hCG is used as the maturation
Evaluation of the cervix
trigger.
Evaluation of the uterine cavity
In our standard protocol for the natural and the minimal stimulation IVF, our
unique GnRH agonist nasal spray is used for triggering oocyte maturation. Evaluation of the endometrial phases
In the minimal stimulation cycle, we administer 50mg of clomiphene citrate Evaluation of the tubal ostia
65 Geneva
The hysteroscopic surgery of pathologies like a submucous fibroids, uterine Proteomics technology enables the evaluation of protein function at the
septum and endometrial polyps, before IVF/ET treatment, improves results molecular level. Testicular spermatogonial stem cells will permit the use of
Prospective studies demonstrates that at present is hysteroscopic surgery is gene-therapy in severest forms of spermatogenesis defects.
the method of choice to improve the cumulative pregnancy rate as well as The use of high magnification light microscopy imaging method is
the live birth rate in selected women with intracavitary pathologies and a complementing classical light microscopy in evaluating sperm morphology.
history of reproductive failure. Fertilization potential is assessed by sperm staining assays using antibodies
Surgical procedures through hysteroscopy must be reserved to experienced against acrosomal proteins and sperm penetration assay.
hysteroscopists. Determination of sperm apoptosis markers, i.e. mitochondrial membrane
potential integrity, plasma membrane translocation of phosphatidylserine,
404.2 – OOCYTE SHARING AS TREATMENT OF FERTILITY caspase activation and DNA denaturation/fragmentation provides
information on sperm function, which assists the clinician with therapy
Kamal Ahuja, The London Women’s Clinic, London, UK. decision.
The introduction of egg sharing in the UK in 1992 was greeted with deep The detection of reactive oxygen species (ROS) deserves to be routinely
suspicion. Despite its practical appeal to patients, the concept was performed. Besides classical luminescence, flow cytometry technology
portrayed as exploitative and unethical and it deeply polarised the opinions permits accurate intracellular ROS determination.
of key stakeholders and also the media. The HFEA came under tremendous
pressure to ban the practice and seemingly came very close to doing just Before ICSI, sperm is selected by swim-up, glass wool filtration or density
that. However, to their credit they resorted to a careful examination of the gradients. The hyaluronic acid binding identifies mature sperm with low
arguments through public debates and a consultation exercise before, in frequency of aneuploidy.
1998, accepting egg sharing as a licensed treatment. In 2007, some ten Magnetic-activated cell sorting using annexin V-conjugated microbeads
years after the original decision, the HFEA reaffirmed their support of the eliminates apoptotic sperm. Sperm selection could also be based on
original decision by approving egg sharing as a suitable practice to obtain membrane electrostatic charge.
human eggs for stem cell research. Sperm selection at high magnification using Nomarski interference contrast
It is probably fair to say that apart from IVF itself no assisted conception is useful to identify more precisely the size and the number of nuclear
procedure has undergone such an in-depth scrutiny as egg sharing before vacuoles.
becoming accepted. The key objectives of this presentation are: 1.To After testicular or epididymal extraction, sperm motility-vitality is enhanced
examine how the HFEA argued the merits of egg sharing and what was by antioxidants.
initially perceived as the shortcomings of egg sharing, 2. The promise egg
sharing may hold for clinical research and treatment in future, and, 3. The Excellent post-thaw motility is being observed after testicular sperm
vigilance and research required to ensure that the practice does not deviate freezing.
from its objective of providing a safe and affordable form of IVF treatment Sperm selection tests will be presented and comments based on the
for those who otherwise face difficulties in procuring IVF treatment. author’s experience provided.
405.1 – OOCYTE MATURATION AND ANEUPLOIDY: THE MOLECULAR 407.1 – STUDY ON EFFECTS OF MALE REPRODUCTIVE TRACT INFECTIONS
PROTAGONIST IS APC WITH AEROBIC BACTERIA ON SPERM QUALITY
Keith T. Jones, The University of Newcastle Australia, Newcastle, Callaghan, A. Kaluarachchi, S.Wijeratne, P.K.B.Mahesh, H.R. Seneviratne. Department
Australia. of Obstetrics & Gynecology, Faculty of Medicine, Colombo, Sri Lanka.
Oocyte quality is a major factor governing a woman’s fertility. Quality is poor Introduction: This study was performed to evaluate the association between
when chromosome segregation errors occur during oocyte maturation. male reproductive tract infections with aerobic bacteria and semen quality
Such errors, which include trisomy 21, affect between 20-40% of all human among men who are seeking infertility treatment at a tertiary care setting.
oocytes, and go on to produce mostly non-viable, aneuploid embryos;
Methodology: Male partners of infertile couples who presented for treatment
making chromosome segregation errors the leading cause of early embryo
to the professorial unit at the De Soyza Hospital for Women were recruited
loss. The aetiology of aneuploidy in maturing oocytes has been heavily
for the study (n=200). Seminal fluid analysis and seminal fluid culture were
investigated but remains obscure, with female age being the only well-
performed according to the WHO criteria. Mid-stream urine culture was
characterized correlate.
performed on the same day to exclude urinary tract infections. Results were
The Spindle Assembly Checkpoint (SAC) is a universal, error-detecting analyzed using Statistical Package for Social Sciences (SPSS) 15.0.
mechanism, employed by all dividing cells to stall cell division until
Results: Normal sperm concentration was seen in 69.5% (n=139/193);
chromosomes are ready to segregate. Although it was first thought that the
normal sperm motility was seen in 39.5 %( n=79/193);normal sperm
SAC would be defective or absent in maturing oocytes, this is now known
morphology was seen in 38% (n=76/179) and normal sperm vitality was
not to be the case. Instead, we have recently found that the maturing
seen in 36.5 %( n=73/179). Pathogenic organisms were isolated in seminal
oocyte’s susceptibility to chromosome mis-segregation is due to its peculiar
fluid in 8% of males (n=16/191) with a colony count of more than 103,out of
and unique regulation of the SAC’s target: the Anaphase-Promoting Complex
which two also had concomitant urinary tract infections with the same
(APC) during oocyte maturation.
pathogen. Another 7.5%(n=15/191) showed a mixed growth of organisms.
Organisms isolated in positive cultures were, Staphylococcus aureus in
406.3 – HOW TO STUDY AND SELECT THE BEST SPERM FOR ICSI 37.5%(n=6), beta(β) hemolytic Streptococci in 18.8%, (n=3), alpha(α)
hemolytic Streptococci in 12.5% (n=2), non-hemolytic Streptococci in 6.3%
Branko Zorn, Andrology Centre, Department of Obstetrics and Gynaecology, (n=1) and Escherichia coli in 5.9% (n=1). Analysis did not show a significant
University Medical Centre Ljubljana, Ljubljana, Slovenia. association between the seminal fluid infections with aerobic organisms and
Since the introduction of intracytoplasmic sperm injection (ICSI) much seminal fluid parameters. P values for individual parameters were,
progress has been made in resolving male infertility due to sperm-oocyte concentration (0.465), motility (0.608), morphology (0.869) and viability
fusion defects. In order to improve the results of ICSI in all causes of male (0.137).
infertility, selection of the best sperm becomes a priority. This has been Conclusion: Infections with aerobic organisms have not shown an effect on
made possible by the development of various methods for sperm evaluation. seminal fluid parameters.
In infertile men, numerous candidate genes and transcripts involved in
sperm cell development and fertilization are being studied using microarray
analysis; they allow the recognition of genetic and epigenetic factors of male
infertility. Sperm aneuploidy screening in sperm from the ejaculate or testis
is recommended for prediction of implantation.
April 19-22, 2009 66

Conclusion(s) Similar to a large uterine septum, a small partial uterine

407.2 – SEMINAL MORPHOLOGY AND THE OUTCOME OF ASSISTED septum or arcuate uterus negatively influences the implantation in
REPRODUCTION TECHNIQUES stimulated IVF and ICSI cycles .
A. Kaluarachchi1; S. Wijeratne1, P.K.B.Mahesh1, C. Nelson2,

H.R. Seneviratne1. 1Department of Obstetrics & Gynecology, Faculty of 407.4 – A NOVEL PROCEDURE FOR SEVERE CASES OF ADENOMYOSIS —
Medicine; 2 Vindana Reproductive Health Centre; Colombo, Sri Lanka. SURGICAL TREATMENT BY THE TRIPLE-FLAP METHOD FOR
Introduction: This study was performed to assess the value of assessing RECONSTRUCTION OF THE UTERINE WALL- OSADA PROCEDURE FOR
sperm morphology and its relationship with the outcome of In Vitro MASSIVE ADENOMYOSIS
Fertilization (IVF) and Intra Cytoplasmic Sperm Injection (ICSI) . Hisao Osada1, Toshiyuki Kakinuma1, Tom Kiyosi Fujii1, Sherman J Silber2,
Material and Methods- A descriptive correlation study was done using the Keiichi Kato3, Osamu Kato3. 1Department of Obstetrics and Gynecology,
records of 306 couples who have sought In Vitro Fertilization (IVF) and Intra Nihon University School of Medicine, Tokyo, Japan 2 Infertility Center of St.
Cytoplasmic Sperm Injection (ICSI) treatment for sub fertility during 2003 to Louis, St. Luke’s Hospital, St. Louis, U.S.A. 3 Kato Ladies Clinic, Tokyo,
2008 at a tertiary care centre. Data was analyzed using Statistical Package Japan.
for Social Sciences (SPSS) 15.0. The male partner’s sperm morphology was Introduction: A multiple flap reconstruction of the uterine wall following
compared with Fertilization Rate, pregnancy and pregnancy outcome. wide excision of adenomyosis tissue was developed for women with severe
Results - Out of the 306 couples (mean age-Female-35.91 Male-38.65), dysmenorrhea and infertility. Consecuctive series of operations in women
238(77.8%) had undergone IVF alone and 49(16.0%) had ICSI, while with severe adenomyosis.
19(6.2%) was treated with both techniques. In 300 (98.03%) of them Materials and Methods: Severe adenomyosis causes infertility, severe
husband was the sperm-provider and a donor provided sperms in dysmenorrhea, and hypermenorrhea. Surgical reconstruction is problematic
6(1.97%).The mean value of normal morphology was 34.39%( SD-13.45%). because of absence of any surgical plane or capsule, and fear of uterine
According to the World Health Organization criteria, 193(63.71%) of the rupture with pregnancy. In fact, the usual treatment is hysterectomy. Our
sperm providers/donors had normal morphology. The mean Fertilization treatment for even the most severe cases of adenomyosis involves wide
Rate was 67.45% (SD-25.6%). The correlation between the morphology and complete excision of affected tissues to assure adequate removal, followed
Fertilization Rate was significant (R=0.030, P=0.004). Mean values of Head, by a multiple flap reconstruction of the uterine wall without overlapping
Mid Piece and Tail abnormalities were 59.86%, 12.91% and 11.59% suture lines to prevent ruptures in subsequent pregnancies.
respectively. Head and Tail abnormalities correlated negatively with the Results: The results were as follows: 1) Wider and more thorough excision
Fertilization Rate, but it was not statistically significant. (Head- of the affected tissues were possible compared to the conventional
R=0.037/P=0.088, Mid Piece-R=0.001/P=0.788, Tail-R=0.24/P=0.171).Out conservative surgery using a wedge resection. 2) The massive tissue
of the 76(24.84%) who became pregnant with IVF and ICSI 20(26.3%) of defects created by the wide excision of the lesion could be reconstructed
them miscarried the pregnancy. There was no significant association with an adequate thickness of uterine wall by overlapping the thin remaining
between successful pregnancy and morphology (P=0.178) or having a myometrial musculature with non-overlapping suture lines. 3) There were
miscarriage and the morphology (P=0.976). no complications such as interstitial haematoma, suture diastasis,
Conclusion- Sperm morphology has a significant impact on fertilization adhesions, etc. observed. 4) There was remarkable improvement in
while the relationship between successful pregnancy and miscarriage was dysmenorrheal and hypermenorrhea. 5) Twenty-six women wished to
not significant. Detail analysis of the type of morphological abnormality is conceive following the surgical removal of the adenomyosis. Sixteen of
useful since it is related to the success of IVF and ICSI.. them (61.5%) subsequently conceived. 13 cases went to term and all were
delivered by elective Caesarean section. There were no cases of uterine
rupture. 6) There was only a 3.84% recurrence (4 cases) of the
407.3 – UTERINE SEPTUM DECREASES THE IMPLANTATION RATE IN IVF / adenomyosis.
ICSI CYCLES
Conclusions: Wide complete excision of adenomyosis with multiflap
Tomaz Tomazevic, Helena Ban-Frangez, Irma,Virant-Klun, Ivan Verdenik. reconstruction of uterine wall results in a dramatic reduction in both
Slovenia, Ljubljana, Slovenia. menstrual cramping and menstrual flow volume post-surgically, and in
Objective: To evaluate the effect of hysteroscopic resection of a large uterine women who desired to conceive, allowed them to go to term without uterine
septum (Class V according to the American Fertility Society (AFS) rupture.
classification) and of a small partial uterine septum (Class VI according to
AFS classification or arcuate uterus) on implantation after ET in stimulated
IVF and ICSI cycles. 407.5 – PREVALENCE OF MARKERS FOR THROMBOPHILIA AND
IMMUNOLOGICAL DISORDERS IN PATIENTS HAVING IN VITRO
Study design: The retrospective matched control study included 339 ETs in FERTILISATION AND EMBRYO TRANSFER
stimulated IVF or ICSI cycles before hysteroscopic resection of a large or
small partial uterine septum (113 and 176 ETs respectively) and 538 ETs in L. Mettler, A. Salmassi, A. G Schmutzler. Department of Obstetrics and
stimulated IVF or ICSI following hysteroscopic resection of a large or small Gynaecology, University of Kiel, Kiel, Germany
partial uterine septum ( 226 and 248 ETs respectively). For each ET in the Background: Birth rates after IVF remain low at 30%, causes are
study group, we found two consecutive ETs s from the IVF/ICSI registry discussable. Investigators reported an increased prevalence of single
who had a normal uterus and were matched for age, BMI, stimulation thrombophilic or immunologic disorders.
protocol, quality of embryos, the use of IVF or ICSI and for various Methods: In a first uncontrolled pilot study we studied 123 patients
infertility causes. The clinical pregnancy rate was the main outcome undergoing IVF. Prior to initiation of IVF, patients were interviewed for
measure. Data on the septum length were obtained during hysteroscopic historical and clinical evidence of hypercoagulatory state or immunological
resection by comparing the length of the 1.3 cm long yellow tip of the disease. Blood samples were determined using a pattern of hereditary
electric knife to the length of the resected septum. (protein-S, -C-deficiency, factor V Leiden), acquired thrombophilia
Results: The implantation rate before hysteroscopic metroplasty was (antiphospholipid antibodies) and immunological parameters (ANA,
significantly lower , both in women with a small partial septum ( 13,6% complement factor 4).
before resection vs. 25,6% in the normal controls, OR 2,9; p<0,001) and a Results: No patient displayed clinical evidence of active immunological or
large septum ( 12,4 % before resection vs. 29,2% in normal controls, OR hypercoagulative disease. Prevalence of protein S deficiency (11.2%),
2,1; p<0,002) compared to women with a normal uterus. After the surgery, positive antiphospholipid antibodies (32.5%) and ANA (30%) were
the pregnancy rate was comparable to the pregnancy rate in women with significantly higher than in the normal population (< 1%; p < 0.005; ~2%; p
normal uterus: in both women with a small partial ( 23,2 % vs. 25,4%) and < 0.005; ~5%; p < 0.005). Overall 82/123 (66.7%) of the patients were
in women with a larger septum ( 21,8% vs. 20,4%) respectively. positive for any pathological marker, 31/123 (25.2%) of the patients had a
combination of parameters of hereditary and/or acquired thrombophilia
67 Geneva
and/or autoantibodies. Women with multiple abnormal tests were found to We summarise ethiopathogeny, characteristic, supervision and treatment of
have a strong tendency towards a lower pregnancy rate after IVF although these complications
the difference did not reach statistical significance (38.7% vs. 51.2% We remind measures to prevent medical and legal complications of OHSS
respectively; p > 0.005).
Conclusions: In an unselected population of patients undergoing IVF, a
substantial share (66.7%) demonstrates pathological assays for 408.1 – INCIDENCE OF ANEUPLOIDIES IN OVARIAN STIMULATION
thrombophilia or immunological disorder without symptoms of clinically TECHNIQUES
active disease. Every fourth patient (25.2%) displays combinations of these J. Remohí, Institut Universitari-Instituto Valenciano de Infertilidad. Valencia.
laboratory parameters. These patients show a strong tendency towards España.
decreased pregnancy rates after IVF. Therefore a risk-adapted strategy in
Several factors affecting gametes and embryos have been found to be
treatment with low dose aspirin, heparin and/or prednisolone to decrease
related to an increase in chromosome abnormalities: changes in
the IVF failure rate seems appropiate.
temperature during oocyte culture and handling (Pickering et al., 1990;
Almeida and Bolton, 1995), ageing of gametes (Badenas et al., 1989; Munné
407.6 – ASSESSMENT OF THE OVARIAN RESERVE FOLLOWING and Estop, 1993), use of a 20% oxygen tension instead of 5% (Pabon et al.,
CHEMOTHERAPY AMONG A COHORT OF CANCER PATIENTS WHO 1989; McKiernan and Bavister, 1990), hormonal stimulation in some mouse
UNDERWENT UNILATERAL OOPHORECTOMY VERSUS PARTIAL strains (Maudlin and Fraser, 1977), sub-optimal stimulation in humans
OOPHORECTOMY FOR OVARIAN TISSUE CRYOPRESERVATION (Hammitt et al., 1993). Furthermore, in patients with high response to
gonadotrophins there was an increased incidence of diploid oocytes (Tarin
Abir Massarwa, Ami Amit, Dalit Ben-Yosef, Tanya Cohen, Tamar Shwartz, et al., 1990) and “in vitro” maturation (IVM) experiments with higher
Nava Mieraz, Rona Limor, Foad Azem. Racine IVF Unit, Lis Maternity concentrations of FSH in the culture media showed increased aneuploidy
Hospital, Sourasky Medical Center, Tel-Aviv, Israel. rates in MII oocytes (Roberts et al., 2005). All these data suggested that
Background: The number of patients surviving cancer is increasing, hence superovulation protocols in IVF might increased the risk of aneuploidy in the
reproductive potential becoming an important issue. Many reports have resulting embryos.
suggested some parameters i.e.: FSH, anti Mulerian hormone and number Previous results from our group (Reis Soares, 2003) showed that, in oocyte
of follicles as for the assessment of ovarian reserve following chemo- donation cycles, those donors with higher response to standard doses of
radiotherapy. gonadotrophins produced significantly higher number of chromosomally
Methods: The study included 24 cancer patients. 12 patients underwent abnormal embryos. Theses results are in concordance with other authors
partial oophorectomty (Group A) and 12 patients underwent unilateral that showed higher incidence of aneuploidy with higher number of oocytes
oophorectomy (Group B). Prior to freezing, the ovarian cortex was dissected retrieved (Munné et al., 2006, Baart et al., 2007).
into 1- to 2-mm and cut into 10 X5-mm sections. Twelve months following As the results were unexpected, we decide to run another study. We selected
the end of chemotherapy we assessed several parameters of ovarian reserve young donors that were classified as “high responders” using a
including: menstrual status, day 3 (basal): FSH, E2, AMH and antral follicle conventional stimulation protocol. These donors had a first cycle with more
count (AFC). In 21 patients all parameters were analyzed while in 3 patients than 20 oocytes and/or serum estradiol levels >3,000 pg/mL, on hCG day,
only AMH was available. without developing ovarian hyperstimulation syndrome We then performed
Results: The two groups were comparable in regard to average age, two consecutive stimulation treatments on these donors. In treatment 1,
indications for chemotherapy, levels of basal FSH, AMH and AFC. We found donors received similar stimulation dose than in the initial cycle: 225 IU/day
positive correlation between FSH, AMH and AFC (p = 0.001) and (p = 0.019) of recombinant follicle-stimulating hormone (r-FSH, Gonal-F ®; Serono,
correspondingly . Five patients in each group had reduced ovarian reserve Madrid, Spain or Puregon; Organon, Madrid, Spain) for the first 5 days,
Conclusions: Ovarian reserve as assessed by the measurement of AMH, when serum E2 was assessed and gonadotrophin dose adjusted if
basal FSH and antral follicles count, showed no statistical difference necessary. In treatment 2, decreased daily doses of gonadotrophins were
between the study groups. The lack of difference is attributed to the fact that administered: 150 IU/day of recombinant follicle-stimulating hormone (r-
the two groups were comparable in the average age, indications for FSH, Gonal-F ®; Serono, Madrid, Spain or Puregon; Organon, Madrid,
cryopreservation and mode of chemo/radiotherapy. In both groups ovaries Spain). Preimplantation Genetic Screening (PGS) was performed on
were exposed to chemotherapeutic agents which affected similarly the embryos resulting from the two stimulation regimens. Chromosomes 13,
ovarian reserve. In cases in which the damage was severe the ovarian 15, 16, 17, 18, 21, 22, X and Y (Vysis Inc. Downers Grove, IL, USA) were
reserve decreased below a threshold independent of the original ovarian analyzed and chromosomally normal embryos were transferred to the
reserve. While in cases in which the chemotherapeutic protocol mildly recipients on day-5. Exclusion criteria for oocyte recipients were: previous
affected the ovary, the ovaries reserve stay above a threshold independent of history of recurrent miscarriages or repetitive implantation failures; severe
the original mass. male factor (<5x106sperm/mL or <5% normal morphology sperm); other
indications for PGS. Our results showed a significant decrease in the mean
In patients in whom the ovarian reserve could severely be affected following number of retrieved oocytes (22.4 vs. 14.9; p=0.0004) as well as in serum
chemotherapy; one should consider application of oophorectmy and E2 levels (3030.4 IU vs. 2085.6; p=0.0025) with the mild-stimulation
cryopreservation whole ovary rather than partial cryopreservation. protocol. The percentage of abnormal embryos was similar in both
protocols (55.9% vs. 50.4%), as well as blastocyst rates (68.4%% vs.
74.8%) in the conventional stimulation compared to mild-stimulation
407.7 – VASCULAR COMPLICATIONS OF OVARIAN HYPERSTIMULATION
protocol. There was no difference between the two stimulation protocols in
(OHS)
the average number of chromosomally normal blastocyst obtained per
J-M. Dreyfus, A.Watrelot, A.Khebbe.; CRES, Lyon, France. donor (2.8±2.2 vs. 2.4±1.7). The clinical efficiency of the donation cycles
Treatment of infertility and A.R.T are now routinely performed. Efficiency of was similar in the conventional and mild-stimulation protocols, in terms of
drugs and specially rFSH, agonists, antagonists in specific protocols the percentage of pregnancies achieved after donation (63.0% vs. 57.9%),
increases success rates. implantation rates (30.2 vs. 31.6) and number of live-births (10 after each
stimulation protocol). A moderate reduction on oocyte number and E2 levels
However, OHS is difficult to control and may conduct to an Ovarian Hyper
was observed in a subgroup of 12 donors. In this subgroup, significantly
Stimulation Syndrome (OHSS).Frequency of complications is probably
higher blastocyst rates were observed in the mild-stimulation protocol
underestimated.
compared to conventional stimulation (78.9%% vs. 61.1%; p=0.0493) and
The goal of this communication is to review vascular (arterial and venous) ongoing pregnancy rates with the mild-stimulation protocol doubled those
complications.Some of them can be particularly grave and possibly fatal. achieved with the conventional protocol (66.7% vs. 33.3%). These
We describe the type, the delay of occurrence, and the issue of the reported differences were translated into 9 live-births after mild stimulation and 5
accidents in the literature. We report two arterial complications recently live-births after conventional stimulation. We concluded that in high
happened in Lyon responder donors, both stimulation regimens (conventional vs. mild-
April 19-22, 2009 68

stimulation) had a similar efficiency in producing chromosomally normal - Verpoest W, Fauser BC, Papanikolaou E, Staessen C, Van Landuyt L,
blastocysts and live-births. Therefore, a moderate decrease in daily Donoso P, Tournaye H, Liebaers I, Devroey P. Chromosomal aneuploidy in
gonadotrophins doses would not compromise the number of embryos conceived with unstimulated cycle IVF. Hum Reprod. 2008
chromosomally normal embryos available for transfer and the take home Oct;23(10):2369-71.
baby rates. Despite inter-donor differences could be expected to this
decrease in gonadotrophins doses.
408.2 – BIOLOGY AND BIOCHEMISTRY OF AMH/MIS
In parallel, we run another study in normosresponse donors to compare
embryo aneuploidies in natural vs. stimulated cycles in the same egg donor. Nathalie Josso, Nathalie di Clemente, Rodolfo Rey, J.Y. Picard. INSERM Unit
Firstly, the donor underwent a non stimulated cycle. When the follicle 782, Clamart, France.
reached 18 mm in diameter, rCG was administered and oocyte retrieval was The existence of a fetal testicular product responsible for the regression of
scheduled 36 h later. PGS was performed in the resulting embryos following Müllerian ducts in male fetuses was discovered by Alfred Jost in the early
a similar laboratory protocol. Secondly, donors followed ovarian stimulation fifties. The “Müllerian inhibitor” as he called it is now known as anti-
under a GnRH agonist long protocol and a starting fixed dose of 150 UI of Müllerian hormone (AMH) or Müllerian inhibiting substance (MIS). A
rFSH and 75 UI of hp-hMG. Recovered oocytes were inseminated with the member of the TGF-ß superfamily, it is produced by Sertoli cells, mainly
same sperm donor. All the embryos from the stimulated cycle were also before puberty, and by postnatal granulosa cells. The level of circulating
analyzed and a maximum of 2 euploid embryos were transferred. Similar AMH, which varies according to sex and developmental stage, provides
aneuploidy rate was observed in the natural vs. the stimulated cycle (44.8% valuable information on the functional activity of gonadal somatic cells. Axel
vs. 48.6%) and therefore, we concluded that ovarian stimulation would not Themmen and Bart Fauser, in Rotterdam, have shown that serum AMH is
increase the incidence of chromosomally abnormal embryos in this group of also an excellent marker of ovarian reserve (1), hence its increasing value in
patients. Pregnancy rate was significantly higher in stimulated cycles the field of assisted reproduction. In contrast, apart from its key role in
compared to natural cycles (50.0% vs. 13.3%; p=0.0006). inducing the regression of Müllerian ducts in male fetuses at 8 fetal weeks,
References its biological action does not appear to significantly affect human gonadal
physiology, in either males or females.
- Almeida PA, Bolton VN. The effect of temperature fluctuations on the
cytoskeletal organisation and chromosomal constitution of the human To induce regression of Müllerian ducts in the human fetus, AMH must be
oocyte. Zygote. 1995; 3 (4): 357-65. expressed before 8 fetal weeks. As shown by Zhan et al (2), coelomic
epithelial cells expressing AMH receptors under the control of Wnt7 (3)
- Baart EB, Martini E, Eijkemans MJ, Van Opstal D, Beckers NG, Verhoeff undergo epithelio-mesenchymal transformation and migrate into the peri-
A,Macklon NS, Fauser BC. Milder ovarian stimulation for in-vitro Müllerian epithelium. AMH action also leads to a wave of apoptosis and to
fertilization reduces aneuploidy in the human preimplantation embryo: a an accumulation of cytoplasmic ß-catenin in the peri-Müllerian mesenchyme
randomized controlled trial. Hum Reprod. 2007; 22 (4):980-8. (4). In the absence of AMH or its receptors, the Müllerian ducts develop into
- Badenas J, Santalo J, Calafell JM, Estop AM, Egozcue J. Effect of the the Fallopian tubes, uterus and upper vagina. Retention of Müllerian duct
degree of maturation of mouse oocytes at fertilization: a source of derivatives occurs in genetic males harboring mutations of AMH or its type
chromosome imbalance. Gamete Res. 1989; 24 (2):205-18. II receptor (5) leading to the persistent Müllerian duct syndrome. In male
- Hammitt DG, Syrop CH, Van Voorhis BJ, Walker DL, Miller TM, Barud KM. rodents, AMH also negatively affects Leydig cell differentiation and function
Maturational asynchrony between oocyte cumulus-coronal morphology (6).
and nuclear maturity in gonadotropin-releasing hormone agonist In female mice, AMH controls the recruitment of primordial follicles into the
stimulations. Fertil Steril. 1993; 59 (2):375-81. growing follicle (see (7) for review). The higher rate of primordial
- Maudlin I, Fraser LR. The effect of PMSG dose on the incidence of recruitment in AMH–deficient transgenic mice leads to premature
chromosomal anomalies in mouse embryos fertilized in vitro. J Reprod exhaustion of the follicle pool causing an early cessation of ovulation in
Fertil. 1977; 50 (2): 275-80. ageing animals. Mice heterozygous for the AMH-null allele have an
intermediate phenotype. Thus mothers of PMDS patients would be expected
- McKiernan SH, Bavister BD. Environmental variables influencing in vitro to undergo early menopause but data are not yet available. AMH also
development of hamster 2-cell embryos to the blastocyst stage. Biol decreases sensitivity to FSH and may inhibit FSH-dependent selection of
Reprod. 1990; 43 (3): 404-13. follicles for dominance.
- Munne S, Estop AM. Chromosome analysis of human spermatozoa stored AMH biochemistry is relatively straightforward. A glycoprotein homodimer
in vitro. Hum Reprod. 1993; 8 (4): 581-6. linked by disulfide bonds, it undergoes proteolytic processing to generate an
- Munne S, Ary J, Zouves C, Escudero T, Barnes F, Cinioglu C, Ary B, Cohen active 25 kDa N-terminal dimer. Processing is required for biological activity,
J. Wide range of chromosome abnormalities in the embryos of young egg as the latter is destroyed by mutations that block cleavage (8). AMH signals
donors. Reprod Biomed Online. 2006; 12 (3): 340-6. through a specific type 2 receptor and through type I receptors shared with
the BMP family, ALK-2 and ALK-3. Smads 1, 5 and 8 act as its cytoplasmic
- Pabon JE Jr, Findley WE, Gibbons WE. The toxic effect of short exposures
effectors (9).
to the atmospheric oxygen concentration on early mouse embryonic
development. Fertil Steril. 1989; 51 (5): 896-900. Reference List
- Pickering SJ, Braude PR, Johnson MH, Cant A, Currie J. Transient cooling 1. vanRooij, I. A. J., Broekmans, F. J. M., teVelde, E. R., Fauser, B. C. J. M.,
to room temperature can cause irreversible disruption of the meiotic Bancsi, L. F. J. M., de Jong, F. H., and Themmen, A. P. N. (2002) Hum
spindle in the human oocyte.Fertil Steril. 1990; 54 (1): 102-8. Reprod 17, 3065-3071
- Reis Soares S, Rubio C, Rodrigo L, Simon C, Remohi J, Pellicer A. High 2. Zhan, Y., Fujino, A., MacLaughlin, D. T., Manganaro, T. F., Szotek, P. P.,
frequency of chromosomal abnormalities in embryos obtained from oocyte Arango, N. A., Teixeira, J., and Donahoe, P. K. (2006) Development 133,
donation cycles. Fertil Steril. 2003; 80 (3): 656-7. 2359-2369
- Roberts R, Iatropoulou A, Ciantar D, Stark J, Becker DL, Franks S, Hardy K. 3. Parr, B. A. and McMahon, A. P. (1998) Nature 395, 707-710
Follicle-stimulating hormone affects metaphase I chromosome alignment 4. Allard, S., Adin, P., Gouédard, L., di Clemente, N., Josso, N., Orgebin-
and increases aneuploidy in mouse oocytes matured in vitro. Biol Reprod. Crist, M. C., Picard, J. Y., and Xavier, F. (2000) Development 127,
2005; 72(1): 107-18. 3349-3360
- Tarin JJ and Pellicer A. Consequences of high ovarian response to 5. Josso, N., Picard, J. Y., Rey, R., and di Clemente, N. (2006) Pediatric
gonadotropins: a cytogenetic analysis of unfertilized human oocytes. Fertil Endocrinology Reviews 3, 347-358
Steril. 1990; 54 (4): 665-70.
6. Racine, C., Rey, R., Forest, M. G., Louis, F., Ferre, A., Huhtaniemi, I.,
- Verberg MF, Macklon NS, Nargund G, Frydman R, Devroey P, Broekmans Josso, N., and di Clemente, N. (1998) Proc. Natl. Acad. Sci. (USA) 95, 594-
FJ, Fauser BC. Mild ovarian stimulation for IVF. Hum Reprod Update. 2009 599
Jan-Feb;15(1):13-29.
69 Geneva
7. Visser, J. A. and Themmen, A. P. N. (2005) Mol Cell Endocrinol 234, 81-86 understand the emotional processes that a patient navigates in accepting
8. Belville, C., Van Vlijmen, H., Ehrenfels, C., Pepinsky, R. B., Rezaie, A. R., ending IVF, as well as being cognizant of factors that predict adaptation so
Picard, J. Y., Josso, N., di Clemente, N., and Cate, R. L. (2004) Mol the needs of the patient are better met at this endpoint. Effective and
Endocrinol 18, 708-721 compassionate doctor-patient communication has been shown to lessen the
emotional sting of ending IVF for couples and contributes to patients be
9. Orvis, G. D., Jamin, S. P., Kwan, K. M., Mishina, Y., Kaartinen, V. M., more open to alternatives such as egg donation.
Huang, S., Roberts, A. B., Umans, L., Huylebroeck, D., Zwijsen, A., Wang,
D., Martin, J. F., and Behringer, R. R. (2008) Biol Reprod 78, 994-1001
410.1 – EFFECTS OF L-GLUTAMINE AND STRAW SIZE, FREEZING RATE AND
THAWING RATE UPON POST-THAW QUALITY OF HUMAN
409.3 – FUNDING IVF TREATMENT - THE CANADIAN EXPERIENCE SPERMATOZOA
Beverly Hanck, Infertility Awareness Association of Canada (IAAC), Hussian Asherkaci, L.M. Aboshala, M.A. Danfour, O.A. Elsraite, M.S.
Montreal, QC, Canada. Elmahaishi. Faculty of Science, Faculty of Medicine, 7th October University
This presentation sets out the rationale for government funding of in vitro and Misurata Infertility Centre, Misurata, Libya.
fertilization (IVF) and other assisted reproductive technologies (ART) in The objective of this study was to investigate cryoprotective effect of L-
Canada, and includes an economic analysis to support the recommendation glutamine, straw size, freezing rate and thawing rate in preserving motility of
that Canada’s public health care system should provide safe and effective human spermatozoa during the freezing-thawing process. MATERIALS AND
infertility treatment for all Canadians who need it. METHODS: Normal Semen sample were collected by ejaculation, according
In Canada, the need for IVF far exceeds its accessibility, and the treatment to criteria of the World Health Organization (WHO), as well as spermatic
remains financially out of reach for many infertile Canadian couples. morphology according to the strict Kruger criterion. Swim up technique was
Because there has been virtually no funding, infertile Canadian couples often carried out to obtain 5-6 X 106 progressive motile sperms with a good
resort to cheaper but less effective alternatives such as ovarian stimulation morphology. In Experiment 1, three straw sizes (1mm, 2mm and 5mm),
and/or hormone injections. These regimens have a major downside in that three freezing rates (straws suspended 2 , 7 and 9 cm above liquid nitrogen)
there is a significant risk of multiple pregnancy. With ovarian stimulation, and three thawing rates (in water at 20, 30 and 37 degrees C) upon post-
poor control over the number of mature eggs produced may result in the thaw quality of sperm, and to determine the best treatment combination.
birth of triplets, quadruplets and even higher-order multiples. Statistics Quality was expressed in terms of the percentage progressively motile
show multiple-pregnancy rates of 30% through ovarian stimulation. sperm 5 min after thawing. In Experiment 2, the best straw size, freezing
rate and thawing rate in preserving motility of human spermatozoa were
In the last ten years, Canada’s birthrate has dropped 25%, while the number
used to investigate the effects of L-glutamine at different concentrations
of multiple births has increased by 25% over the same period. Available data
(20mM and 50mM) on post-thaw sperm motility. Data were analyzed by
from Europe and North America suggest that infertility treatment accounts
means of a repeated measures factorial analysis of variance and means
for 30% to 50% of all twin births and for up to 80% of all higher-order
compared. Results: There were significant effects by straw size, freezing
multiple births. Of these higher-order multiple births about 50% are
rate, thawing rate on human sperm cryopreservation procedure (P<0.05).
attributable to fertility pills and hormone injections.
The straw size 2mm, straws suspended above liquid nitrogen 7cm and
Multiple pregnancies have very broad repercussions: they severely affect thawing rates 37 degrees C were the optimal conditions for preservation of
families psychologically, medically and financially, while the cost to human spermatozoa. Using these conditions we found that addition of L-
Canadian provinces is far greater than for singleton pregnancies, due to glutamine with 50mM improve the survival rate of human sperm. In
increased needs for medical and social support. Multiple pregnancies also conclusion: These new preservation protocol permit extended conservation
lead to elevated health risks for mothers and infants, increased perinatal and of viable spermatozoa that may capable of supporting normal embryonic
neonatal costs and, in extreme cases, lifelong costs because of the development and the live birth of healthy baby after ICSI.
disabilities that occur more frequently in multiples and multiple related pre-
term births.
410.2 – IN VITRO FERTILIZATION IN NATURAL CYCLE FOR CONCURRENT
Accordingly the authors recommend the following strategies: (1) education
TRANSFER OF FRESH AND FROZEN/THAWED EMBRYOS
of the medical and allied professions, as well as prospective parents; (2)
К.Ilyin. IVF & Genetics Centre “FertiMed”, Moscow, Russia.

monitoring of women during ovarian stimulation treatments, with the option M. Anshina, A. Smirnova, N. Shamugia, E. Abliaeva, T. Troshina, I Kalinina,
to switch to IVF; (3) reduction of the number of embryos transferred in IVF
treatment and encouragement of elective single embryo transfer (eSET), Introduction: In women with normal ovulation we usually perform the
with couples seeking treatment at younger ages; (4) optimal availability of transfer of cryopreserved embryo(s) in natural cycle. The key point is
IVF treatment, based on a 100% refundable tax credit, the cost of which appropriate timing of ovulation with consequent blastocyst thawing and
would be easily offset by significant savings associated with reduced transfer on day 5 after ovulation. Pregnancy rate is high enough – 31% per
incidence of multiple pregnancies. embryo transfer, but we suggested that simultaneous transfer of
These strategies should result in at least a 50% decrease in the rate of cryopreserved embryo(s) and fresh embryo from the natural cycle may
multiple births in every province that adopts funded IVF. improve the results.
Objective: to identify and compare the clinical pregnancy rate and multiple
pregnancy rate after the transfer of cryopreserved embryos and after
409.4 – PSYCHOLOGICAL COMPLEXITIES SURROUNDING ENDING IVF
concurrent transfer of fresh embryo obtained after IVF in natural cycle and
TREATMENT
cryopreserved embryos.
Janet Takefman, McGill Reproductive Centre, McGill University Health
Material and Methods: Two groups of women who had not more than 3
Centre, Montreal, QC, Canada.
cryopreserved embryos at blastocyste after previous IVF or IVF/ICSI cycles:
A question frequently posed by infertility specialists is when to stop IVF group I – 47 women who underwent 55 transfers of cryopreserved embryos
treatment or when to advise patients that “enough is enough”. The answer in natural cycle; group II – 29 women who underwent 31 natural cycle
to this question is challenging for a number of reasons. First, there is always follicle aspirations and concurrent fresh/frozen embryo transfers. In all
a theoretical probability of success with further attempts. Second, there is patients follicle aspiration was performed 26-32 hours after hCG injection
no continuum of benefits with each successive attempt. Finally, the (5000 IU). All embryos were transferred on day 5 after ovulation (group I) or
physician and patient likely base their decision to end IVF on different follicle aspiration (group II).
criteria. The physician will consider medical and risk factors, the patient,
Results: In group I, 55 embryo transfers resulted in 17 pregnancies: 14
financial, personal and emotional costs. Telling a couple that IVF is no longer
singleton and 3 multiple. Two pregnancies stopped to develop at 6 and 8
recommended usually implies their dream of having a genetic child will be
weeks of gestation. In group II, 31 embryo transfers resulted in 19
unrealized. Research has documented this transition process for couples as
pregnancies: 11 singleton and 8 twins. Totally 79 embryos were transferred
being long, difficult and complex. It is important that the physician
in group II: 31 fresh embryos (9 at 6-8-cells stage and 22 at blastocyste
April 19-22, 2009 70

stage) and 48 frozen/thawed blastocystes. There were 3 twins after the synergic effect of FSH and / or LH with other nutrients required for folliclular
transfer of one fresh and one frozen embryos. Spontaneous reduction of development in long - term culture.
one fetus at 6-8 weeks of gestation occurred in three of eight twin
pregnancies and in one singleton pregnancy.
410.4 – VITRIFICATION OF DAY 3 EMBRYOS IMPROVES THE POST THAW
There was no difference in women’s age, mean number of transferred PREGNANCY RATES
embryos and endometrium thickness on the day of transfer between groups.
The clinical pregnancy rates per transfer was higher in group II in compare Hrishikesh Pai, Nandita Palshetkar, Rishma Pai. Lilavati Hospital IVF Centre,
to group I (61% vs 31%, P=0,002). The implantation rate (34% vs 17%, Lilavati Hospital, Bandra, Mumbai, India.
P=0,04) and multiple pregnancy rate (42% vs 14%, P=0,01) also were Introduction: Vitrification improves the oocyte, pronuclear stage and
higher in group II. blastocyst freezing survival. However there are very few studies which have
Conclusions: Concurrent fresh/frozen embryo transfer is preferable reported application of vitrification to 6-8 cell human embryos on day 3 of
especially in patients with small number of frozen embryos. It may improve culture. . In this study we describe the technique as well as demonstrate the
clinical pregnancy rate and reduce the rate of transfer cancellation when significantly favorable outcome of day 3 vitrified embryos.
embryos failed to survive after thawing. Only in three patients we are Material and Methods :Since January 2007 , vitrification of day 3 embryos
confident that both fresh and frozen/thawed embryos were implanted. But was carried out with the open technique of Cryotop (kitazato,japan ) using
high multiple pregnancy rate possibly indicates high implantation potency of two step protocol with ethylene glycol, DMSO and sucrose as
both types of transferred embryos. cryoprotectants.After equilibration in vitrification media, 1 to 2 embryos
were loaded on to the tip of the Cryotop straw in a minimum volume of <0.1
microlitre. . The straw was then plunged into liquid nitrogen. The tip was
410.3 – N-ACETYL-CYSTEIN IMPROVES RESULTS OF LONG-TERM CULTURE covered with cover straw and stored. Patients were prepared for frozen thaw
OF FROZEN / THAWED HUMAN OVARIAN TISSUE embryo transfer using depot Gnrh agonist in conjunction with hormone
R Fabbri1, G. Pasquinelli2, L. Montanaro3, V. Magnani1, F. Tamburini1, D. replacement therapy. An average of 2 to 3 Embryos was thawed using a four
Keane4, Y. Cabello Vives5, S. Venturoli1. 1Human Reproductive Medicine Unit, step protocol. The embryo survival in the form of > 50 % blastomeres intact
S. Orsola-Malpighi Hospital, University of Bologna, Bologna, Italy; 2Clinical and 100 % blastomere intact was calculated. Assisted laser hatching in the
Pathology Section, Department of Radiological and Histocytopathological form of partial zona thinning was carried out on all embryos .After 2 hours
Sciences, S.Orsola-Malpighi Hospital and 3Department of Experimental of culture in the incubator, the embryo transfer was carried out.
Pathology, University of Bologna, Bologna, Italy; 4Department of Obstetrics Results: 220 patients were subjected to vitrification. 70 frozen thaw
and Gynaecology, Royal College of Surgeons in Ireland, Rotunda Hospital, vitrification cycles were done from January 2007 till date. 95 % of embryos
Dublin, Ireland; 5Human Assisted Reproduction Laboratory, FIV thawed had 100% intact blastomeres. The B HCG positive rate was nearly
RECOLETOS, Policlinico Nuestra Senora de America, Madrid, Spain. 68.5 % (48/70) and the clinical pregnancy rate was 61.4(43/70) % per
Introduction: Women treated for malignant disease have a risk of premature embryo transfer.
ovarian failure (POF). Their fertility may be preserved by ovarian tissue culture in Conclusion: This is one of the few studies on day 3 vitrification of 6 to 8 cell
vitro. With in-vitro culture, ovarian tissue is usually kept under normoxic embryos. The method is significantly successful. However the largest
conditions; the oxygen concentration being considerably higher than in-vivo. drawback in the open method is the potential risk of contaminating the
Thus, the oxidative stress, the excessive production of reactive oxygen specimen straws .Ongoing trials comparing the outcome of vitrified embryos using a
(ROS) may contribute to the atresia observed in in-vitro cultures. The aim of our closed system and an open system are being carried out in our unit.
study was to investigate the effect of N-Acetyl-Cystein (NAC), a radical-
scavenger, supplemented to culture media on follicular and stromal preservation.
Material and Methods: The ovarian cortex of 3 patients was collected and 410.5 – HIGH SURVIVAL AND PREGNANCY RATE AFTER SINGLE EMBRYO
immediately cryopreserved, as previously described, Fabbri et al, 2003. After TRANSFER USING CRYOTOP BLASTOCYST VITRIFICATION METHOD
thawing, samples of ovarian cortex from each patient were collected for Tsuyoshi Okubo, Shoukichi Teramoto, Masashige,Kuwayama. Shinbashi
molecular and histological analysis (control t0). Additional samples were cultured Yume Clinic, Shinbashi Minato-ku, Tokyo, Japan.
at 37°C in an atmosphere of 6% CO2 for 16 weeks, in minimum essential
Objective: Multiple pregnancy is one of the most serious problem in IVF-ET,
medium (α-MEM) supplemented with antibiotics, Follicle Stimulating Hormone
but single embryo transfer (SET) can solve the problem. Various bad
(FSH), Insulin-Transferrin-Selenium (ITS), Human Serum (HS), with NAC(Medium
influences considered, SET is highly recommended to pregnant patient in
A) or without NAC (Medium B), or ITS, HS, FSH and Luteinizing Hormone (LH) with
case of IVF-ET. Since the opening of our clinic, we have cryopreserved all of
a peak level every 28 days, with NAC (Medium C) or without NAC (Medium D).
embryos by the cryotop vitrification method in substitute for the slow
The medium was changed every second day. Sample collection for morphological freezing method. That survival rate was almost 95%(n=1089), this
and molecular analyses was carried out after 8 (t8) and 16 (t16) weeks of culture. surprisingly good result shows that almost of all vitrified blastocysts can be
The number and developmental stage of follicles was determined using Light transferred with very little damage. We have performed SET by using
Microscopy (LM) on semithin sections stained with toluidine blue; follicular and cryotop blastocyst vitrification method. Then we report our clinical outcome
stromal cell integrity was evaluated using Transmission Electron Microscopy that is high survival and pregnancy rate but no multiple pregnancies.
(TEM) and assessment of the expression of GDF-9, Bcl-2 and Bax mRNAs by
Design: The study in our clinic was conducted from May 2007 to August
Real Time RT-PCR.
2008 for 695 patients in as the minimal stimulation IVF cycles. A total of
Results: LM and TEM showed an improved follicular and stromal structural 695 patients were transferred that one cryopreserved blastocyst vitrified by
preservation in Medium A with respect to Medium B. Molecular evaluation the cryotop vitrification method.
demonstrated Medium A had increased expression of GDF-9 transcripts and a
Materials and Methods: All of the patients were treated by the minimal
high Bcl-2/Bax ratio; these results were consistent with the good follicular and
stimulation IVF cycles using clomiphene citrate and recombinant FSH.
stromal preservation observed by morphological analyses in medium A with
Administration of 50 mg clomiphene citrate was initiated on cycle day 3 and
respect to medium B.
that of 150 IU recombinant FSH was injected every other day from day 8.
The beneficial NAC effect was also apparent when comparing Medium C with D. When the size of the dominant follicle and the estradiol concentration
However in this experimental condition the Bcl2/Bax ratio was apparently not reached the predefined values, gonadotropin-releasing hormone agonist was
affected. administered to induce follicular maturation. Oocytes were then retrieved at
Conclusions: These results suggest that NAC is necessary to perform long-term aspiration 3035 h later since dosed nasal GnRHa spray. Then matured
cultures of human ovarian cortical tissue kept under normoxic conditions.The oocytes were inseminated by conventional IVF or ICSI procedure. When the
beneficial NAC effect is most likely related to its intrinsic radical-scavenger blastocyst developed over 170μm diameter, they were vitrified by the
activity which minimizes the excessive production of ROS, one of the major cryotop vitrification method (Kuwayama, 2006) on day 5 to 7. All blastocyst
determinants of human ovarian cortical tissue degeneration in long-term were underwent laser assisted hatching after thawing, then it was cultured
cultures. Reducing ROS tissue damage with NAC is necessary to maximize the for before ET at least for 2 hours retrieval. Blastocysts were transferred to
71 Geneva
the patient visibility of the catheter under ultrasound guide.

Results: The mean number of oocytes retrieved was 2.8 per treatment cycle. 410.7 – FROZEN EMBRYO TRANSFER: THE EFFECT OF NUMBER
The number of patients cryopreserved blastocysts by cryotop vitrification TRANSFERRED, STAGE TRANSFERRED AND SYNCHRONISATION ON
method was 695 and mean age of patient was 37.0±4.1 years old. Of all THE CLINICAL OUTCOME
1029 IVF-ET cycles initiated, the rate for blastocyst survival on day 5, 6 and Tarique Salman, Ariel Zosmer, Amanda,Tozer, Luca Sabatini, Talha Al-
7 were 99.0%(489/494), 91.2%(486/533) and 85.5%(53/62), respectively. Shawaf. Barts and the London Centre of Reproductive Medicine, Rochdale,
The rates for clinical pregnancy, ongoing pregnancy and multiple pregnancy Lancs, UK.
were 50.9%(524/1029), 43.8%(451/1029) and 0%(0/1029) , respectively.
Introduction: Elective single embryo transfer in fresh and in frozen embryo
Conclusions: The Cryotop vitrification method enabled us to obtain excellent transfer (FET) cycles has been advocated to reduce multiple pregnancies. In
blastocyst survival rate on day 5 to 7. Furthermore the SET protocol using this study we analysed the effect of transferring 1 vs. 2 good quality
in this study resulted in highly pregnancy rate without multiple pregnancies. embryos, the number of embryos thawed, the number of blastomeres at
In short, the Cryotop vitrification blastocysts method is very valuable for freezing and the synchronisation of the embryos and the endometrial
SET, it can be possible for patients both maintaining high pregnancy rate development at transfer on the outcome of FETs.
and preventing from multiple pregnancies under an appropriate endometrial
condition. Material and Methods: We retrospectively analysed 818 FET cycles. 48
(5.6%) cycles resulted in failed thaw, leaving 770 consecutive non donor
oocytes FET cycles. Good quality embryo was defined as <15%
410.6 – COMPARISON OF VITRIFICATION VERSUS SLOW COOLING FOR fragmentation post thaw. The transfer of day 2 (D2) embryos on day 3 (D3)
BLASTOCYST CRYOPRESERVATION IN AN IVF PROGRAM post ovulation (in natural cycle FETs) or after the start of progesterone
Andrew Dorfmann, Micheal Geltinger, Michael,Sisson, Simone Yap, Iwaszko supplementation (in HRT cycle FETs) or D3 embryos transferred on D2
Melissa. Genetics & IVF Institute, Fairfax, VA, USA. respectively was defined as an asynchronous FET cycle.
Introduction: In recent years vitrification has been utilized as an alternative Results: The overall clinical pregnancy (CPR) and live birth (LBR) rates were
to conventional slow cooling methods for the cryopreservation of embryos 18.3% and 14.3% per transfer respectively. One embryo was transferred in
and oocytes. In the present study either slow cooling or closed vitrification 123 (15.9%), 2 embryos were transferred in 632 (82.1%) and 3 embryos
(Cryotip™) was used to cryopreserve blastocysts. The purpose of this trial were transferred in 8 (1%) cycles. In 413 (53.6%) of the FET cycles
was to determine if we could improve our results using a closed vitrification embryos of good quality were only transferred, in the remainder the
system for cryopreservation of blastocysts. Results were compared between embryos quality were mixed.
a standard slow cooling method and vitrification. Analysis of outcome by the number of good embryos transferred: One good
Materials and Methods: Embryo culture was done by conventional quality embryo (1xFET) was transferred in 74 cycles, resulting in 6 (8.1 %)
methods using Vitrolife G series sequential media. All embryos were CPR and LBR. Two good quality embryos (2xFET) were transferred in 339
cultured in 5% O2 and 6% CO2. All embryos were cultured to the blastocyst cycles resulting in 83 (24.7%) CPR and 68 confirmed LBR (20.1%) (8
stage prior to cryopreservation on day 5 or 6. Embryos were only selected if pregnancies final outcome is unknown). The CPR and LBR were significantly
they had a well developed inner cell mass, a clear and healthy higher in the 2xFET vs. 1xFET (p=0.015).
trophectoderm layer, and a blastocoel cavity comprising at least 50% of the Twin sacs were noticed in 21 cases and triplets in 2. Twelve (10.3% per
mass of the embryo. Vitrification was carried out using Cryotips® from LBR) sets of twins, and 2 (1.7% per LBR) set of triplets were delivered. The
Irvine scientific using previously published methodology. Slow cooling was overall multiple LBR was 12%.
performed using 0.25 cc straws and subsequently thawed with thaw kits Analysis of outcome by the number of embryos thawed: Cycles where only
from Vitrolife. Statistical comparisons were performed using chi-squared 1 embryo was transferred the LBR was 1.9% when only 1 embryo was
analysis. thawed, 9% when 2 embryos were thawed and 14.2% when more than 2
Results: During the study period, June 2007 through December 2008:167 embryos were thawed. In cycles where 2 embryos were transferred CPR and
patient thaws were attempted; 82 patient thaws using their own oocytes of LBR were 20.5% and 17.6% when only 2 embryos were thawed, 17.1% and
which 29 were slow cooled and 53 were vitrified and 85 patients using 12.8% when 3 embryos were thawed and 22.4% and 19.4% when more
donor oocytes of which 33 were slow cooled and 52 were vitrified. 464 total than 3 embryos were thawed respectively.
Blastocysts were thawed or warmed; 280 were vitrified, and 184 were frozen Analysis of outcome by the number of cells at cryopreservation: When 2
by slow cooling; 225 from patients own oocytes and 239 from donor embryos were transferred and both were frozen at the 2 cells stage the LBR
oocytes. The overall survival rate for slow cooled blastocysts was 83/184 was 5.1% but if both embryos had 3 or more cells when frozen the LBR was
(45%) vs 206/280 (73.5 %). For patients using their own eggs the survival significantly higher at 20.2% (p =0.0014 ).
rate was 34/88 (39%) in the slow cool group and 102/138 from the
vitrification group (74%). From patients using donor oocytes the survival Effect of synchronising freezing day to transfer day: Out of 770 cycles, 94
rate was 49/97 (50%) and 104/142 (73%) respectively. Differences between (12.5%) were asynchronous by +/- 1 day. There was no statistical difference
all groups were statistically significant; p<0.001. The clinical ongoing or in the CPR (18.5% vs. 17.0%) or LBR (14.9% vs. 15.9%) between the
delivered pregnancy rates were as follows: patients using their own oocytes; synchronous and asynchronous groups respectively.
slow cooling: 7/29 (24%) versus vitrification 22/52 (42%) (Not statistically Conclusions: Transferring 2 good quality embryos in FET cycles resulted in
significant), patients using donor oocytes; slow cooling 9/33 (27%) vs significantly higher CP and LB than single FET. The transfer of embryos
vitrification 26/52 (51%) (p < 0.05). frozen at 3 or more blastomeres stage is more successful than that of
Conclusions: Vitrification is a highly efficient and successful method of embryos frozen at 2 blastomeres stage. One day asynchronisation between
cryopreservation when used for blastocyst stage embryos. In our hands it embryos’ and endometrial development does not affect LBR.
yielded significantly better results than that achieved with standard slow
cooling methodologies. Embryo survival, viability, and quality were excellent
and produced good pregnancy rates in this initial series.
April 19-22, 2009 72

ABSTRACTS – POSTERS
revealed that many of the mitochondria of morulae and blastocysts cultured

P.001 – MISOPROSTOL FOR RIPENING OF CERVIX IN NULLIGRAVIDAS in serum-supplemented medium were immature, consistent with a
HYSTEROSCOPY correlation between respiration activity and development of mitochondria.
Seddigheh Abdollahi-Fard, Abolfazl Bohlouli. Tabriz University of Medical Table 1: Oxygen consumption rates of the bovine embryos cultured in
Sciences, Tabriz, Eastern Azarbaijan, Iran. serum-free medium (IVD101) and serum-supplemented medium
(HPM199+CS)
Hysteroscopy is an operation in which the gynecologist examines the uterine
cavity using a small telescope inserted via the vagina and the cervix and this Embryonic stage Oxygen consumption rate (×1014/mol•s-1)
instrument is to diagnostic and operative purpose in intrauterine pathology. IVD101 HPM199+CS
In the non pregnant state the cervical canal is very narrow and the cervix 2 cell 0.46±0.05 (17) 0.52±0.04 (6)
resists mechanical force to open it and there is much complication inherent
to mechanical dilatation including cervical tearing, uterine perforation, and 4 cell 0.45±0.03 (17) 0.47±0.04 (6)
bleeding and intra abdominal organs damage. these complications is more 8 cell 0.46±0.02 (10) 0.52±0.04 (10)
profound in nulliparous patients.
Morula 1.10±0.05 (23) 0.70±0.05 (12) *
Material and Methods: To determine the effect of self administered oral
Blastocyst 1.99±0.07 (35) 1.33±0.10 (12) *
misoprostol 200 mg (a synthetic prostaglandin E1 analogue) vaginally the
night before procedure to all infertile or sub fertile patients with *Significant differences (P<0.05, compared with embryos cultured in
hysteroscopy indication and compared with non used cases.108 patients IVD101 at same embryonic stages). The numbers in parentheses represent
with indication for hysteroscopy including vaginal bleeding, septate uterus, the numbers of embryos examined.
endometrial thickness or mass (polyp,submucosal myoma) were included Conclusion: These results demonstrated that the culture conditions (serum
and half of patients offered self administered misoprostol 200 mg the night supplemented or not) affect the respiratory activity, mitochondrial
before admission and the other half did not get any medication before morphology, and embryo quality. Measuring oxygen consumption with
procedure for cervix ripening .the procedure time and complications and SECM may have broad applications for determining suitable culture
surgeon satisfaction was compared in two groups. conditions for embryos.
Results: the effect of misoprostol in ripening of the cervix was significant
even in nulliparous post menopause patients and the hegar no-7dilatator
P.003 – ENDOMETRIOSIS SEVERITY AND IVF OUTCOME
was replaced without any resistance.
Germain Agnani, Arnaud Collin, Xavier Dellis, Pascale Lagré, Christine
Conclusion: We would recommend this inexpensive and easy to use regimen
Souquet, Christophe Roux, Christianne Joanne, Didier Riethmuller. Service
to infertile or sub fertile women prior to undergoing operative hysteroscopy
de Gynécologie Obstétrique, CHU Besançon, Besançon, France.
to reduce the risk of complications and facilitate cervical dilatation.
Objective: The aim of this study was to evaluate the impact of endometriosis
on In Vitro Fertilization (IVF) outcome, focusing on the initial surgical
P.002 – RESPIRATORY ACTIVITY AND ULTRASTRUCTURAL FEATURES OF approach, surgical difficulties, and ovarian reserve.
BOVINE EMBRYOS DEVELOPED IN DIFFERENT CULTURE SYSTEMS
Retrospective review of 200 consecutive first oocyte retrievals in patients
USING SERUM-FREE OR SERUM-CONTAINING MEDIA
with endometriosis.
Hiroyuki Abe1, Shoko Yamashita2, Hiroyoshi Hoshi2. 1Graduate School of
Materials and methods: Severity of endometriosis was evaluated during the
Science and Engineering, Yamagata University, Yonezawa; 2Research
first laparoscopy according to the American Fertility Society (AFS)
Institute for the Functional Peptides, Yamagata; Japan.
classification and a simplified classification
Introduction: Scanning electrochemical microscopy (SECM) measuring
( 3 subgroups).
system has been employed to quantify the respiratory activity of embryos in
several animal species including humans. Respiration is a useful parameter Subgroup A : superficial peritoneal and ovarian implants.
for evaluating embryo quality as it provides important information about Subgroup B : endometrioma (diameter ≥ 5mm) without deeply infiltrating
metabolic activity. Recently, we have found that there is correlation between endometriosis.
embryo quality and respiratory activity in bovine embryos. It has been
reported that culture condition affects embryo quality and serum may be a Subgroup C : deeply infiltrating endometriosis involving the bladder,
key factor. The aims of this study were: (1) to assess the oxygen sigmoid, uterosacral ligaments or vagina.
consumption; (2) to examine the ultrastructural features of bovine embryos Male infertility and anovulation were excluded. Surgical treatment was
at different developmental stages cultured in serum-free and serum- always performed before IVF.
supplemented media. Follicular count was estimated just before the first IVF attempt.
Materials and Methods: Bovine oocytes were matured in IVMD101 medium Data were analysed using the chi square test for qualitative variables and the
(Research Institute for the Functional Peptides: IFP, Japan) and inseminated Kruskal Wallis one-way analysis of variance for quantitative variables,
in BO-based medium. For serum-free culture, inseminated ooocytes were statistical significance was set at p<0.05.
cultured to blastocyst stage in IVD101 medium (IFP, Japan) in an
atmosphere of a low oxygen condition (5% CO2/5% O2/90% N2) at 38.5C˚. Results: Clinical pregnancy rate was significantly lower in group C ( group A
For serum-supplemented culture, inseminated oocytes were cultured in : 42%, group B : 50%, group C : 5.9%, p<0.001 ).
HPM199 medium (IFP, Japan) supplemented with 5% calf serum Subgroups A, B, and C were comparable when we considered female age,
(HPM199+CS) in the presence of bovine cumulus/granulose cells in a BMI, duration of infertility, sperm concentration, basal FSH level,
humidified atmosphere of 5% CO2 in air. Oxygen consumption by individual endometrioma diameter before laparoscopy (groups B & C), follicular count,
bovine embryos was non-invasively quantified by SECM measuring system. oocyte number, embryo number, and classical parameters of stimulation.
Some embryos were prepared for transmission electron microscopy. The percentage of laparotomies and ovarian cystectomies was similar in
Results: Oxygen consumption has been monitored at various developmental subgroups B and C.
stages of bovine embryos cultured in IVD101 and HPM199+CS media (Table Total top quality embryo number and top quality transferred embryo number
1). Oxygen consumption rates of the single embryos were low from 2-cell to were significantly lower in group C (p<0.05).
8-cell stages (0.48-0.52×1014/mol•s-1). In serum-free culture, an increase in
oxygen consumption rate was found at the morula (1.03×1014/mol•s-1) stage Early complications after IVF gradually increased with the severity of
and blastocysts showed an even higher oxygen consumption rate endometriosis ( group A : 5%, group B : 20%, group C : 38%, p<0.001 ).
(1.86×1014/mol•s-1). On the other hand, the oxygen consumption of morulae The surgical approach including a second look offered the best results
and blastocysts produced in serum-supplemented medium were lower than (p<0.01).
those of embryos cultured in serum-free medium. Ultrastructural analysis
Clinical pregnancy rate was also significantly influenced by total embryo
73 Geneva
number (p<0.01), top quality embryo number, top quality transferred

embryo number (p<0.001), detection of endometrioma before stimulation P.005 – IMPACT OF SURGICAL TREATMENT OF VARICOCELE ON THE
(p<0.02), and BMI (p<0.02). BIOLOGICAL ASSESSEMENT OF IN VITRO FERTILIZATION (IVF)
In a multivariate logistic regression including top quality transferred embryo Mounir Ajina, Loussaief Wafa, Ibala Samira, Ben Regaya Lassad, Hidar
number, endometriosis subgroups, BMI, and also age, basal FSH level, and Samir, Khairi Hedi, Saad Ali. Unit of Medicine of Reproduction, Sousse,
follicular count, three variables were selected : top quality transferred Tunisia.
embryo number, endometriosis subgroups, and detection of an
endometrioma. Fewer differences were observed when the AFS classification Introduction: Surgical treatment of varicocele makes it possible to restore
was considered. the adequate temperature of spermatogenesis, but the re-establishment of
the fertilizing capacity remains doubtful. The aim of this work is to evaluate
Conclusions: As opposed to recent studies which minimize the impact of the impact of varicocele and of its surgical repair on the biological
endometriosis on IVF results, our study suggested that the persistence of assessment of in vitro fertilization.
active lesions is more deleterious than mild follicular destruction and
adhesions. Materials and Methods: We studied 97 patients carrying varicocele, 46% of
which was operated. Among these patients, 16% had a grade III and 55%
had the grade II. Varicocele was on the left in 79% of case. All the couples
P.004 – HOW DO HYPOGONADOTROPHIC HYPOGONADISM PATIENTS DO IN profited at least from a cycle of in vitro fertilization and for each couple we
ASSISTED REPRODUCTION CYCLES? analyzed: semen parameters, fertilization rate, segmentation rate and
Gülnaz Şahin, Cüneyt Barut, Ayşin Akdoğan, Halil Abdülrezzak, Rafael Levi, embryonic quality. Statistical study was made by software SPSS11.0.
Ege Tavmergen Göker, Erol Tavmergen. Ege University Family Planning and Results: Average age of patients was of 38±5, 08 years with a primary
Infertility Center, Izmir, Turkey. infertility duration average of 6, 81±3, 95 years. Infertility was of male origin
Introduction: Hypogonadotrophic hypogonadism (HH) is a rare condition of in 55% of case and mixed origin in 40%. The analysis of semen finds: a
female infertility. The purpose of this study was to evaluate the assisted mean spermatic concentration of 23, 05±35,71millions/ml, a mean mobility
reproduction treatment (ART) cycle characteristics and outcomes of women (a+b) of 15±10%, % of abnormal forms of 65±30% and 30±28 % of dead
with HH and compare them with a group of normal responders’ cycle spermatozoa. 12% of our patients were azoospermic with an average rate of
outcomes. FSH of 7, 28±4, 78 UI/ml. assisted fertilization was indicated in 68% of
couples and traditional fertilization in 11%of couples. During the first cycle,
Design: Retrospective analysis 7, 77±4, 93 follicles were aspirated of which only 66, 48±33, 09% reached
Materials and Methods: Analysis was carried out on 26 HH female patients the stage of zygote after insemination. The fertilization and segmentation
with 40 ART/ICSI cycles during 1996-2008 performed at Ege University rates were respectively of 50, 96±35, 81% and 81, 02±103, 49%. The
Family Planning and Infertility Center. Age matched 66 healthy women were average number of type I embryo transferred was of 1, 82±1, 25.
selected as a control group with a history of regular menses, normal basal Conclusion: The biological assessment of in vitro fertilization was faded
endocrine hormone levels and undergone ART with long protocol cycles for among patients carrying varicocele, especially the fertilization rate. On the
male factor infertility from the same center. Controlled ovarian stimulation hand segmentation rate and embryonic quality were both preserved. In our
was performed with human menopausal gonadotrophin (HMG) in HH group study the surgical clean of varicocele does not improve the oocyte
and recombinant FSH combined with gonadotrophin releasing hormone fertilization rate. This same rate does not vary also to a significant degree
agonist for control group. according to the grade of varicocele.
All IVF cycle outcomes were evaluated. Their age, infertility duration, basal
serum hormone levels, the mean diameters of both ovaries on
ultrasonographic examination, total and daily dosage of gonadotrophins, P.006 – NONENZYMATIC ANTIOXYDANT SEMINAL ANALYSIS IN INFERTILE
serum E2 levels and LH levels on the day of human chorionic gonadotrophin MEN AND ITS EFFECT ON THE BIOLOGICAL ASSESSEMENT OF IN
(HCG) injection, number of total oocytes, metaphase II (MII) oocytes, VITRO FERTILIZATION (IVF)
transferred embryos, implantation rates and pregnancy rates were analyzed. Mounir Ajina1, Atig Fatma1, Ibala Samira1, Loussaef Wafa1, Hidar Samir1,
Statistical analysis was performed with SPSS version 15.0. Mann-Whitney - Khairi Hedi1, Kerkeni Abdelhamid2, Saad Ali1. 1Unit of Medicine of the
U test, Chi-square tests were used to compare groups. p < 0.05 was Reproduction, Hospital F. Hached; 2Laboratory of Biophysics, Faculty of
considered statistically significant. Medicine, H Bourguiba Monastir; Sousse, Tunisia.
Results: Totally 36 cycles in HH group and 59 cycles in control group Introduction: The aim of this study consists on evaluating the nonenzymatic
reached to the embryo transfer. The mean ages in the HH group were antioxidant statute of infertile patients programmed for in vitro fertilization
33±4.5 (range 26-42) and 31.1±2.6 (range 28-37) of the control group. The and the repercussions of this statute on the biological assessment of in vitro
mean infertility durations were 9±3.9 years and 8.4±4.0 years, respectively. fertilization.
The mean diameters of both ovaries on ultrasonographic examination were Material and Methods: Our work concerns 57 couples including six
smaller than control group (p<0.01). Basal serum FSH, LH levels were Witness patients profited from a spermatic nonenzymatic exploration: Zinc,
significantly lower than control group (p<0.05). Although HMG was used in Malondialdéhyde and Glutathion. The proportioning of seminal zinc was
the HH group, the control used only FSH containing preparates. Daily carried out by the spectrophotometry of atomic absorption to flame, the acid
dosage of gonadotrophin requirement and total dosage of gonadotrophins Malondialdéhyde (MDA) was evaluated by the thiobarbituric method of the
were significantly higher in HH patients than controls, 5.6±1.1 vs. 3.7±0.5 acid (TBA) and the proportioning of the various forms of glutathion (GSH)
and 60.7±18.5 vs.31.8±8.6 ampoules, respectively (p<0.01). Duration of (total, oxidized and reduced) was based on the coupling of the groupings
stimulation was significantly longer in HH group (p<0.01). Serum E2 levels thiols of the GSH with the dithiobisnitrobenzoate (DTNB) which is
on the day of HCG injection was not different between groups but mean transformed into a compound coloured in yellow and detectable with 412
serum LH levels on the same day was significantly higher in control group nm.
than HH patients, 2.3±1.5 vs. 0.76± 0.73, respectively (p<0.05). Mean
number of oocytes, MII oocytes were the same. Although more embryos Results: Our results showed that the fertilization rate increased moderately
were transferred in HH group, their implantation rates (15.7±22.8 among patients group. Therefore, oxidative parameters (Zinc, GSH and
vs.25.4±30.5), pregnancy rates per embryo transfer (44.4% vs.55.9%) and MDA) were also raised in patients group than in witness one. In
clinical pregnancy rates per embryo transfer cycle (38.9% vs. 50.8%) asthenozoospermic group of patients, the MDA rate was conversely
seemed to be lower than controls, even though not statistically significant. correlated with mobility (p=0, 04). However, total GSH was positively
correlated with fertilization (p=0, 03) and with embryonic segmentation rate
Conclusions: Although higher consumption of gonadotrophin dosages and (p=0, 02).
longer duration of ovarian stimulation is needed in HH patients, their
pregnancy rates in IVF cycles were not different than normal responder Conclusion : The increase in the seminal concentrations in MDA among
patients. infertile patients can partly explain the bad spermatic quality observed as
well as the weak fertilisation rate. But, the positive correlation of
April 19-22, 2009 74

antioxydants with the rate fertilization confirm the paternal nonenzymatic

antioxydant contribution in the early embryonic development. P.009 – PREDICTORS OF SUCCESS IN GNRH ANTAGONISTS CYCLES FOR IVF-
ICSI
P.007 – OVARIAN HYPERSTIMULATION USING GONADOTROPIN UPREGULATE Aysin Akdogan, Gülnaz Sahin, Cuneyt Barut, Rafael Levi, Ege Goker, Erol
CFTR EXPRESSION IN VIVO: IMPLICATIONS FOR HYDROSALPINX Tavmergen. Family Planning & Infertility Research & Treatment Center, Ege
ENLARGEMENT AND UTERINE FLUID ACCUMULATION DURING University, Izmir, Turkey.
OVULATION INDUCTION Introduction: Controlled ovarian stimulation with GnRH antagonists for
Louis Chukwuemeka Ajonuma1, 2, Lai Ling Tsang1, Sun Yee Lam1, Hsiao prevention of LH surge is largely used in mild ovarian stimulation protocols.
Chang Chan1. 1Epithelial Cell Biology Research Center, Department of The purpose of the study was to evaluate prognostic factors for achieving
Physiology, Faculty of Medicine, The Chinese University of Hong Kong, pregnancy in antagonist cycles.
Shatin, NT, Hong Kong; 2Faculty of Dentistry, The University of Hong Kong, Material & Methods: A total of 1396 IVF patients with their first cycles were
Hong Kong SAR. treated with rec FSH and GnRH antagonist stimulation for IVF –ICSI between
Introduction: Controlled ovarian hyperstimulation (COH) for ovulation 2005-2007 at the Ege University Family Planning and Infertility Research
induction is associated with formation and/or sudden increase in and Treatment Center were evaluated. Patients age, duration of infertility,
hydrosalpinx fluid (HF) as well as reflux into the uterine cavity leading to fertility cause, menstruel regularity, basal endometrium thickness, basal
poor in vitro fertilization (IVF) treatment outcome. However, the mechanisms endocrine parameters (basal FSH, LH, E2, prolactine),total FSH doses and
underlying fluid formation during gonadotropin administration for ovarian dose regulation, total antagonist doses, duration of stimulation, maximum
stimulation have not been thoroughly investigated. Since cystic fibrosis E2, maximum endometrial thickness, number of aspirated oocytes, MII
transmembrane conductance regulator (CFTR), a cAMP-dependent ion oocytes and transferred embryos were recorded for analysis to predict
channel is known to modulate fluid secretion in the female reproductive pregnancy rates. Multivariable logistic regresion analysis was performed
tract, the present study investigated whether ovarian hyperstimulation with a backward LR elimination procedure, a p-value<0,1 was used as a
increases CFTR expression in vivo after gonadotropin administration. criterion for exclusion.
Materials and Methods: We examined CFTR expression after gonadotropin Results: Of the 1396 initiated cycles, 179(12,5%) were cancelled due to:
(HMG, PMSG and HCG) administration in comparison with controls using insufficient responce, empty follicule, fertilisation defect and bad qualty
reverse transcriptase-polymerase chain reaction (RT-PCR), embryos. The pregnancy rate was 40.8%. Over all patients’ mean age was
immunohistochemistry and immunofluorescence staining in a rat ovulation 33,6±5,0 and duration of infertility was 8,7±5,4 years. Infertility causes were
induction model. male factor (52,4%), idiopathic (27,5%), tuboperitoneal factor (10,1%),
Results: RT-PCR revealed that CFTR expression in the uteri of gonadotropin ovulatuar factor (8,6%) and others (1,3%). In a multivariable logistic
treated rats was significantly higher than controls and no difference was regression analysis, age, basal endometrial thickness, basal prolactine
noted in the ovriectomized and control rats. Immunostaining showed levels, maximum endometrial thickness and number of transferred embryos
enhanced CFTR immunoreactivity in the uterine epithelium of rats with intact were found to be associated with pregnancy. The P-values and odds ratios
ovaries when compared to ovriectomized rats. (95%CIs) were respectively 0.042: 0.966(0.934,0.999) for age, 0.013:
0.899(0.826,0.978) for basal endometrium, 0.043: 0.986(0.972, 1.000) for
Conclusions: These results suggest that ovulation induction upregulate the basal prolactine, 0.001: 0.1.153(1.068,1.244) for maximum endometrial
expression of CFTR via ovarian hormones. Upregulation of CFTR leading to thickness, 0.001:1.751(1.394,2.200) for number of transferred embryos.
increased transepithelial fluid transport results in the enlargement of
hydrosalpinx and HF reflux into the uterine cavity during ovarian Conclusions: In this analysis the results revealed that while an increase in
hyperstimulation. These findings may provide grounds for a better treatment women age, basal endometrial thickness and basal prolactine levels cause a
strategy for infertile patients undergoing ART. decrease in pregnancy rates. An increase in maximum endometrial thickness
and number of transferred embryos have a positive effect in pregnancy
rates.
P.008 – BEGIN OF HUMAN LIFE AND RELIGIOUS IDEAS ABOUT RESEARCH ON
HUMAN EMBRYO
P.010 – IMPLANTATION AND CLINICAL PREGNANCY RATES IN DAY 2-3
Mahzad Akbarpour2, Reza Omani Samani1, Seyed Taha Merghati1. TRANSFER VERSUS BLASTOCYST TRANSFER-A RETROSPECTIVE
1
Department of Epidemiology and Reproductive Health, Reproductive COMPARATIVE STUDY
Medicine and Cell Science Research Center; 2Department of Embryology,
Reproductive Medicine and Cell Science Research Center; Royan Institute, H. Tijani, S. Patwardhan, S. Keay, S. Montgomery, R. Kennedy, Rubina Ali.
Tehran, Iran. Centre of Reproductive Medicine, University Hospitals Coventry and
Warwickshire, Coventry, UK.
Doing research on human embryo is a big controversy in around the world
and according to different religions, there are different ideas about it. Begin Introduction: Recent advances in cell culture media have led to a shift in In
of In-vitro fertilization and entrance of embryo donation made a complex vitro fertilisation (IVF) practice from early cleavage embryo transfer to
situation about using human embryos in research. Embryonic stem cells blastocyst stage transfer. Initially, there were contradictory reports that
and hope of treatment of incurable disease made it even more complex. Iran blastocyst transfer did not improve pregnancy rate. However, more recent
is the only Islamic country that practices donation programs and also has evidence has shown significant difference in pregnancy and live birth rates
human embryonic stem cell lines. Here in this paper, we present the Islamic in favour of blastocyst transfer. The purpose of this study was to compare
idea about the begin of human life, permission to do the research on human the reproductive outcome of early cleavage embryo transfer and blastocyst
embryo and therapeutic abortion. The basic presentable conclusions are as stage transfer.
follows: Material and Methods: A retrospective analysis of implantation and Clinical
1. Pre implantation embryo is not considered as human or potential human pregnancy rates was carried out for women who underwent In vitro
and can be used in research with the permission of the parents. fertilization and Intracytoplasmic sperm injection (ICSI) at the Department of
Reproductive Medicine, UHCW, from June 2006 to June 2008. A total of
2. After implantation, although the embryo is not considered human, but 1016 women were involved in the analysis. Of these 524 had IVF while 491
nobody can touch it. Any manipulation of embryo in the uterus is had ICSI. Eight hundred sixty seven had early cleavage embryo transfer
considered try to abort the child. while 149 had blastocyst transfers.
3. After 120 days (for Shiaa Muslims) and 50 days (for Sunni Muslims), it is Results: Evidence of a significant difference in number of positive
believed that soul goes inside the fetus so it is considered a human, and pregnancies between the two treatment groups was detected in favour of
no abortion is allowed. blastocyst transfers: 32.5% in day 2-3 embryo and 38.9% in blastocyst
4. Before this date, it is OK to do the therapeutic abortion if there is an group with unadjusted success rate RR of 1.22, 95% CI of 0.99 to 1.52 and
absolute medical reason. adjusted success rate RR of 1.35, 95% CI of 1.07 to 1.70. A similar trend
75 Geneva
was observed in the successful clinical pregnancies. A total of 28.3% clinical 9.06 oocytes were retrieved and 10.5 ± 6.36 oocytes or 5.25 ± 4.57
pregnancies in day 2-3 embryo group and 34.2% clinical pregnancies in the pronucleids were cryopreserved per patient.
blastocyst group with unadjusted success rate RR of 1.21, 95% CI of 0.95 Time between surgery and chemotherapy was 34.5 ± 9.25 days for patients
to 1.55 and adjusted success rate RR of 1.34, 95% CI of 1.03 to 1.75. with breast cancer undergoing IVF(Proposition: Chemotherapy was not
Adjustment was made for age, FSH, gravidity, number of embryos for and delayed in breast cancer patients undergoing fertility perservation
number of attempts at transfer as well as treatment type, that is IVF or ICSI. techniques with an interval between surgery and adjuvant chemo of 34.5 ±
Conclusions: This study provides evidence that there is a significant 9.25 days.) Peak E2 levels at the time of ovulation was 1210 pmol/L (range
difference in number of positive pregnancies and clinical pregnancies in 540-2190 pmol /L) in patients with breast cancer using the letrozole-
favour of blastocyst transfer. This trend in the difference persists even after gonatrophin- GnRh antagonist protocol and 5550 pmol/L(range 1830-10430
adjusting for confounding factors. This further corroborates recent evidence pmol/L) in patients with other cancers or autoimmune diseases.
in favour of blastocyst transfer in selected patients. Conclusions: Cancer and Fertility is an emerging new field of reproductive
medicine. The approach to patient-care should be multidisciplinary to
P.011 – DEBATE IN EMBRYO DONATION: SOURCE OF EMBRYOS, FROZEN ensure that the best options are offered to women based on current
FROM INFERTILE OR FRESH FROM FERTILE? knowledge whilst ensuring that optimally timed curative oncological
treatment is not compromised. which needs to be approached in the context
Leila Alizadeh, Mohammad Reza Rezania Moalem, Seyed Taha Merghati, of a multidisciplinary strategy for assuring that the best options are offered
Reza Omani Samani. Department of Ethics of Royan Institute, Tehran, Iran. to cancer patients at the right time. In doing this, utmost care should be
For many infertile couples there is no treatment except joining a third party deployed for not disturbing the cancer treatment and notably, delaying the
to the family. Iran is the only Islamic country in which, donation and start of adjuvant chemotherapy.
surrogacy is practiced and in other Islamic countries they are considered
forbidden or “Haram”. Recently, embryo donation law has been approved by
Iran’s parliament and now it is legal in Iran. But there is a misunderstanding
in the source of embryos: they can be obtained from surplus frozen P.013 – THE EFFECTS OF LIF AND EGF ON MOUSE OOCYTE MATURATION AND
embryos of infertile couples or embryos can be made from donated sperms PREIMPLANTATION EMBRYO DEVELOPMENT IN VITRO
and eggs from fertile married couples. Here in this paper we discussed Iraj Amiri1, Ali Amini2, Maryam Parvini2, Narges Mirahadi. 1Dept of Anatomy
ethical, religious and legal aspects of these two procedures and present the and Embroyology, Medical School, Hamadan University of Medical Sciences,
advantages and disadvantages of them. Meanwhile, the new term “both Hamadan; 2Dept. of Biology, Faculty of Science, Kermanshah University,
gamete donation” has been defined for the procedure that is practiced here Kermanshah, Iran.
instead of “embryo donation”. In conclusion we can say: 1) Iranian law
means both “embryo donation” so covers surplus embryos from other Introduction: Recent studies have demonstrated that mammalian
infertile couples and “both gamete donation” so covers making embryos preimplantation embryos are exposed to a mixture of many different growth
from a fertile couple. 2) As gamete donation is practiced in Iran upon factors and cytokines, expressed by the follicles, oviducts and endometrium.
decrees of clergy leaders, we have no law or legislation against “both Receptors for many of these growth factors have also been shown to be
gamete donation”. 3) There are so many ethical, legal and religious expressed by preimplantation embryos. In vitro culture of human and
questions about “both gamete donation” to be answered. 4) Ethical and animal’s embryos in conventional media lacking growth factors can result in
religious questions are very fewer about “embryo donation” comparing to suboptimal growth and a variety of short-term and long-term developmental
“both gamete donation” program, and 5) Embryo sharing is a good way for abnormalities. One of these factors is Leukemia inhibitory factor (LIF).The
donation of fresh embryos. aim of this study was to evaluate the effects of LIF on the mouse
preimplantation embryo development.
Materials and Methods: Six to eight weeks old NMRI mice were
P.012 – MULTIDISCIPLINARY TASK FORCE FOR MANAGEMENT OF WOMEN superovulated by injection of 10IU PMSG and 10IU HCG 48h later. The
WITH CANCER AND FERTILITY PRESERVING ISSUES IN mated mice were killed 48 hours after hCG injection, oviducts were flushed
SWITZERLAND and two-cell embryos collected and divided randomly to two groups
Alexandra Ambrosetti, Marina Bellavia, Victoria Ibecheole, Khalil Zamman, (Control and treatment).Control medium was HTF and treatment medium
Jean-Bernard Dubuisson, Dorothea Wunder-Galié, D. de Ziegler. Onco- was HTF+1000u/ml LIF. In each group the embryos were cultured in an
gynécologie, HUG, Geneva, Suisse. incubator at 37°C with 5% CO2 for 72h.The state of embryo development
Introduction: The desire for future pregnancy expressed by cancer patients was evaluated in 24 hours interval using inverted microscope.
of reproductive age needs to be addressed with a degree of urgency that is Results: There was not any significant difference in the rate of morolla
dictated by the cancer treatment. formation after 48 hours. In comparing blastocyst formation and hatching
The inherent complexity of this emerging field mandates having a rates, 60 and 72 hours after culture, there was significant difference
multidisciplinary structure capable of offering (a-remove) better and safer between control and treatment groups (p< 0.008).
management than individual initiatives might do. We are reporting (We Conclusion: LIF doesn’t provide obvious stimulation in the early mouse
report) the activity of such a multidisciplinary regional task force that exists embryo development until morolla stage; however, it has positive effects on
in Switzerland since 2006, “Réseau Romand de Cancer et Fertilité” (RRCF). blastocyst formation and hatching.
Material and Methods: Between 07/2006 to 01/2009, a total of 32 women Key Words: LIF, Preimplantation embryo, Mouse
(20 breast cancers and 12 other cancers or autoimmune diseases) were
prospectively evaluated for fertility preservation measures before adjuvant
chemotherapy. Of these, 16 underwent controlled ovarian hyperstimulation P.014 – CHROSOMAL ANALYSIS OF THE ABORTUSES IN INFERTILE PATIENTS
(COH) for cryopreservation of 2-PN embryos or oocytes (11 received AFTER TREATMENT
letrozole- gonadotropin-GnRH antagonist protocols, 2 long GnRH-a Tomoka Aniya1, Yoshiharu Nakaoka1, Sachiyo Tarui1, Aya Ohgaki1, Kengo
protocols, 2 GnRH antagonist protocols and 1 a GnRH agonist microflare-up Sugihara1, Mamoru Ida1, Aisaku Fukuda1, Yoshiharu Morimoto2. 1IVF Osaka
protocol). Only 1 woman underwent ovarian tissue cryopreservation. One Clinic; 2IVF Namba Clinic; Osaka, Japan.
woman with squamous cervical cancers (FIGO stages Ia1) underwent
Objectives: Cytogenetic abnormalities of the conceptus are well recognized
trachelectomy. The remaining 14 patients declined any fertility-preserving
causes of pregnancy loss. Most of these abnormalities are the result of
procedure after thorough counselling.
chromosomal accidents at the time of parental meiosis, fertilization and
Results: In women with breast cancer (mean age 31.81, range 25-39), (an early cell division in the zygote. On the other hand, it is well known that
average of) 14 ± 8.73 oocytes were retrieved and 11.66 ± 7.37 oocytes or spontaneous miscarriage rate in infertile patients is approximately 20-25%
7.25 ± 5.80 pronucleids were cryopreserved per patient. In women with and higher than that in natural pregnancy.
other cancers or autoimmune diseases (mean age 30, range 21-39), 11.8 ±
April 19-22, 2009 76

The first objective of the present study was to assess the chromosomal Results: The survival rate in the pronuclear micro injected oocyte using a
abnormality rate by analyzing karyotype of the abortuses in the pregnancies pipette inner diameter 15 or 25 μm were 93% (37 / 40) and 50% (20 / 40)
after infertility treatments. The second objective was to clarify the (P<0.01), and the pronuclear microinjection success rate were 78% (29 /
relationship between chromosomal abnormality rate and maternal age or 37) and 70% (14 / 20), respectively.
early fetal development during pregnancy. Conclusion: The pronuclear that was removed from the oocyte without
Methods: Chromosomal analysis was performed on 289 abortuses from the cytoplasm is able to inject into the enucleated cytoplasm directly. The result
infertile patients. Eighty five cases were conceived by classical infertility shows the possibility that this pronuclear microinjection method becomes
treatment such as timed intercourse or intrauterine insemination, 40 cases the effective treatment of the infertility cause of defective cytoplasm, and of
by conventional IVF (IVF) and 164 cases by intracytoplasmic sperm injection the prevention of heredity of the mitochondrial diseases.
(ICSI). The average maternal age in these 3 groups were not different.
Chorionic villi obtained from dilatation and curettage were cultured and
karyotyped by G-banding techniques. All procedures were performed after P.016 – SWITCHING FROM PN SLOW FREEZING TO BLASTOCYST
obtaining informed consent from the patients. VITRIFICATION: IMPACT ON IVF OUTCOME
Results: Two hundreds seventeen cases (75.0%) had abnormal Marina Argyrou, Massia Moschopoulou, Chris Karamalegos, Sofia Doriza,
chromosome constitutions, in which 152 cases (70.0%) were autosomal Tania Karagianni, Christina Mentorou, Stephen Davies, Minas Mastrominas;
trisomy, 32 cases (14.7%) were structural abnormalities, 8 cases (3.7%) Embryogenesis Assisted Conception Unit, Marousi, Athens, Greece.
were triploidy, 4 cases (1.8%) were tetraploidy and 10 cases (4.6%) were X Introduction: For many years our policy was to routinely slow-freeze
monosomy. Chromosome 16 and chromosome 22 were the most frequent embryos at 2PN stage. In our experience, survival & pregnancy rates after
autosomal trisomies. There are no difference of incidence of chromosomal thawing were better than cleavage stages. We experimented with several
abnormalities in the groups of classical infertility treatment (81.0%), IVF protocols of vitrification, but were not satisfied with survival quality until we
(74.3%) and ICSI (70.8%). But polyploidy rate in either IVF (0%) or ICSI were introduced to the Cryotop method. Vitrification as a procedure
(1.7%) was lower than that in classical infertility treatment group (14.0%). theoretically reduces intracellular damage due to ice crystallization induced
Chromosomal abnormality rate (81.7%) in the advanced maternal age ( 35 by other freezing protocols.
years) was significantly higher than that (63.9%) in younger age (<35
We changed our policy of freezing during July 2008, where patients
years). Aneuploidy rate of chromosomal abnormalities was increased when
undergoing treatment cycles were advised to culture all embryos until
maternal age advanced,
proposed transfer dates and spare blastocyst(s) of good quality vitrified for
Conclusions: The present study had that abortuses in infertile patients after subsequent use as opposed to blindly freezing a proportion of embryos at
any infertility treatment showed high incidence of abnormal karyotype, 2PN stage and allowing the remainder to progress for fresh transfer.
approximately 70 . Assisted reproductive technologies does not only
We assess here the impact of this change in policy had on the outcome of
increase the rate of chromosomal abnormalities in abortuses, but also
IVF cycles during this period of transition.
prevent the polyploidy by observation of number of pronucleus in the
laboratory. The incidence of chromosomal abnormality is strongly related to Materials & Methods: In this study we included all patients from period of
maternal age. February-June 2008 who underwent slow freezing at 2PN stages (Group A,
n=163 patients). They all followed our standard procedure where if 7 or
more fertilized oocytes are available they are advised to have at least 3
P.015 – DEVELOPMENT OF THE SAFETY CYTOPLASMIC EXCHANGE METHOD embryos frozen, & allow the remaining to proceed for transfer either on day
IN PRONUCLEAR STAGE OOCYTES; PREVENTION OF MITOCHONDRIA 3 or at the blastocyst stage, depending upon doctors recommendation.
COEXISTENCE IN CYTOPLASMIC THAT USED THE PRONUCLEAR
The vitrification protocol included in this study were performed between
MICRO INJECTION
July- December 2008 (Group B, n=108 patients). The protocol required that
Fumihito Aono1, Kojiro Kawano1, Masashige Kuwayama1, Yuji Takehara2, all embryos are cultured to the blastocyst stage. Either one or two
Osamu Kato2; 1Advanced Medical Research Institute of Fertility, Kato Ladies blastocysts were transferred, and the remaining blastocysts immediately
Clinic, Shinjuku; 2 Kato Ladies Clinic, Shinjuku; Tokyo, Japan. vitrified using Croptop method.
Introduction: The effectiveness of oocyte cytoplasmic exchange has already Both groups A & B consisted of patients undergoing 1st cycle at our center,
been demonstrated for the treatment of the infertility cause of defective ICSI procedure used, & >6 embryos available. Cycle outcome was then
cytoplasm and of the prevention of heredity of the mitochondrial diseases assessed as clinical pregnancy & implantation rates.
(Kuwayama, 58th ASRM meeting, 2002 . However, mitochondria is brought
Results: Groups A & B did not differ in terms of mean age (Group A:
in with the nuclear when the cell membrane fusion method (McGrath and
34.0±4.2, & Group B: 33.6±3.9), causes of infertility, number of oocytes
Solter, Science 1983) is used, and two kinds of mitochondria, derivation
retrieved (Group A: 14.9±5.5 & Group B: 15.2±5.2 respectively), & number
from both of donor and recipient mitochondria, exist together
of oocytes fertilized (Group A: 8.8±3.4, & Group B: 9.1±3.2). There were
consequentially in the donor cytoplasm. Because mitochondria coexistence
statistical differences in the numbers of embryos transferred (Group A:
does not exist in nature, and the safety in the situation is uncertain, these
2.8±0.2, & Group B: 1.4±0.2), clinical pregnancy rates (Group A:
two types of mitochondria coexistence is an obstruction in a clinical use of
72/163:44.2%, & Group B: 59/108:54.6%) & implantation rates (Group A:
this technology application. To prevent mitochondria coexistence, we tried
104/508:20.5%, & Group B: 71/158:44.9%)
the development of the pronuclear microinjection method.
Conclusions: The switch from routinely freezing at 2PN to Blastocyst had a
Materials and Methods: The pronuclear stage oocytes with informed
fundamental benefit on our IVF programme by improving clinical pregnancy
consent were used for the experiment. A pronuclear was removed from the
rates in 1st cycle patients after transferring fewer embryos. These increased
cell membrane crushed oocyte that was crushed by the pulse of the Piezo
rates are attributed to the patients with blastocyst vitrification having all of
micromanipulator (the cell membrane crushing method) in modified P1
their cohort of embryos cultured, and greater numbers from which to
medium containing 60% of serum substitute supplement (SSS, Irvine
choose the best embryo(s) for ET. Routinely in our programme, we do not
Scientific). To remove the cytoplasm containing mitochondria, the removed
see significant differences in pregnancy rates between day3 & Blastocyst ET.
pronuclear was washed by the repetitive pipetting with the small glass
pipette assembled in the micromanipulator. The cytoplasm-free pronuclear We were allowed to make this change in protocol because of the efficiency
was injected directly to the donor cytoplasm with the glass pipette of the and effectiveness of the cryotop vitrification technique which allows freezing
inside diameter 15 or 25 μm. The pronuclear injected oocytes were cultured & warming blastocysts with virtually no loss of embryos & potential.
with Cleavage Medium (SAGE) containing 10% of SSS for 30 minutes 5%
CO2 5% O2 and 90% N2 at 37C. The oocyte that the cell membrane had
been maintained intact after 30 minutes from the pronuclear microinjection
was judged survival and the oocyte that was able to confirm the pronuclear
intact was judged the pronuclear microinjection success.
77 Geneva
P.017 – EFFECT OF HYDROSTATIC PRESSURE ON CUMULUS AND OOCYTE P.019 – COMPARATIVELY EVALUATION OF INFERTILE (RIF) AND FERTILE
COMPLEX (COCS) DERIVED FROM IN VITRO CULTURED PREANTRAL WOMEN ENDOMETRIAL BIOPSIES AT THE ULTRASTRUCTURAL LEVEL
FOLLICLES BY TEM
Zahra Rashidi, Mehri Azadbakht, Ali Amini. Razi University, Kermanshah, L. Bahar1, Z.N. Candan2, S. Kahraman2, T. Baykal1. 1Mersin University,
Iran. Medicine Faculty, Histology and Embryology Department, Mersin; 2Istanbul
Introduction: In vitro maturation of oocytes is a safe and effective treatment Memorial Hospital, ART & Reproductive Genetics Center, Istanbul; Turkey.
offered in some fertility centers for assisted reproduction. Hydrostatic Introduction: Implantation of embryo in the uterus wall, which involves
pressure as physical force is effective in reproduction system and there is an complex series of interactions and events, is the first requirement for
increase in intrafollicular pressure between 15-20 mmHg in the ovulating embryo to develop beyond the blastocyst stage. Failure in this regard is still
follicle during the late stage of the ovulatory process. Cumulus cells play a widely considered as an obstacle staying ahead of the improvement of the
critical role in oocyte maturation and fertilization. Whether the degree of cell success in assisted reproduction (IVF) and could be mostly attributed to the
death in the cumulus–oocyte complex (COCs) has an impact on oocyte poor endometrial receptivity. During menstrual cycle, endometrium, in
development potential is unclear. Physical forces may to relation to the establishment of endometrial receptivity, undergoes several
assure the incidence of cell death in (COCs). In this study, we examined the morphological changes at the physiological, structural, biochemical and
effect of hydrostatic pressure on the viability of COCs derived from in vitro molecular levels to prepare supportive environment for blastocyst
cultured preantral follicles. acceptance and implantation. Understanding of these changes would
facilitate to discover the still unknown biological mechanisms of
Materials & Methods: Preantral follicles were isolated from 12-day-old implantation. Since problems in pregnancy are frequently associated with
female NMRI mice, each follicle cultured individually in microdrops of MEM- pre and peri-implantation period, considerable advances following the
culture medium under oil during 12 days. On day 12, follicles with extensive research will contribute to the enhancement in both implantation
diameter nearly 500 μm and good quality were induced using 7.5 mIU/ml and pregnancy rates. In this study, we aimed to comparatively investigate
human chorionic gonadotropin (HCG) for in vitro maturation. Then follicles the endometrium tissues of infertile patients with at least 3 previous IVF
were subjected to 20 mmHg hydrostatic pressure for 30 min and cultured failure and fertile patients on the implantation window by transmission
for 24 h. Viability of cumulus cells and oocyte were assessed with electron microscope (TEM) in order to evaluate the differences at the cellular
differential staining (propidium iodide & bisbenzimide) on 0 and 24 h after level.
culture.
Methods: Two groups were defined. In a study group, 21 infertile women,
Results: indicate that, hydrostatic pressure had not changed viability of having history of repeated implantation failure (RIF) in ≥ 3 IVF trails, were
oocyte in 0 and 24 h after culture (p<0.05).At the 0 h, viability of the involved. In a control group, 9 fertile women, who applied to clinic for
cumulus cells were reduced in hydrostatic pressure treated follicles (93%) gynecologic problems, were recruited after having informed consents.
compared to control (97%); (p<0.05). After 24 h viability of the cumulus Endometrial sampling of each patient in both groups was performed on the
cells were reduced in hydrostatic treated follicles (93%) compared to control 19-21 day of the menstrual cycle, which was detected by ultrasonographic
(85%); (p<0.05). examination of existence of corpus luteum and measurement of
Conclusion: hydrostatic pressure had the mild effect on cell death incidence progesterone level. Subsequently, endometrial samples were prepared for
in cumulus cells without any effect on oocyte viability. Hydrostatic pressure TEM analysis and ultra thin sections of randomly selected 3 patients from
may play a role in oocyte maturation and fertilization by improving in each group were comparatively analyzed.
releasing and mediating signals to oocyte. Results: By TEM analysis, all fertile group endometrial biopsies showed
uniformly distributed higher number of pinopodes presenting epithelial cells.
Moreover, less number of ciliated cells among the pinopodes was observed
P.018 – EFFECT OF HYDROSTATIC PRESSURE ON IN VITRO MATURATION OF
as well as cell displaying microvilli at the apical surface were well organized
OOCYTES DERIVED FROM IN VITRO CULTURED PREANTRAL FOLLICLES
with well defined cellular junctions. On the other hand, comparison between
Zahra Rashidi, Mehri Azadbakht, Ali Amini. Razi University, Kermanshah, endometrial biopsies of infertile group and fertile group demonstrated
Iran. marked differences, which could affect the functionality of endometrium and
Introduction: Oocyte in vitro maturation (IVM) is an important reproductive implantation. Less number of pinopodes and rather unmature pinopodes
technology that involves artificial removal of cumulus-oocyte complex were obvious in RIF cases endometrial biopsies. Additionally, more ciliated
(COCs) from antral follicles significant progress has been made in the cells, which were unexpected in the mid luteal phase were still present in
development for the IVM, but there is limited efficiency. Hydrostatic infertile group endometrium and apical surface was less organized with
pressure as physical force is effective in reproduction system and there is an loosen interactions between cell displayed microvilli as well as less number
increase in intrafollicular pressure (15-20 mmHg) in the ovulating follicle of stromal cells with inadequate extracellular cell matrix were observed.
during the late stage of the ovulatory process. In this study, we examined Decidualisation of stromal cells was not frequent and epithelial cells, making
the effect of hydrostatic pressure on in vitro maturation of oocyte derived up endocrine gland were less in number, inadequately vacuolated and
from in vitro cultured preantral follicles. contained prominently heterochromatin nucleus.
Materials and Methods: Preantral follicles were isolated from 12-day-old Discussion: Recurrent implantation failure in IVF is still one of the major
female NMRI mice, each follicle cultured individually in microdrops of MEM- problems remains to be unsolved. In general, underlying causes for RIF are
α culture medium under oil during 12 days. On day 12, follicles with attributed to the problems related with embryos, the endometrial factors or
diameter nearly 500 μm and good quality was induced using 7.5 mIU/ml immunity. However, endometrial based problems during the implantation
human chorionic gonadotropin (HCG) for in vitro maturation. Then follicles window are widely proposed as a main reason for RIF. Despite the
divided in control and experiment groups. In experiment group follicles were mysterious complex interactions between embryo and endometrium during
subjected to 20 mmHg hydrostatic pressure for 30 min and then follicles implantation, comparison of the ultrastructural features of endometrium at
from two groups were cultured for 24-48 h. the implantation window between infertile and fertile patients can deduce
new explanations for RIF. TEM analysis of endometrial samples from RIF
Results: After 24 h percent of metaphase II (MII) oocyte were increased in
group reveals that dramatic changes at the ultrasructural level could be
hydrostatic pressure treated follicles(16%) compared to control (9%);
underlying cause for infertility of these patients. As our best knowledge, this
(p<0.05). After 48 h percent of metaphase II (MII) oocyte were increased in
study was the first comparatively evaluating endometrium of RIF and fertile
hydrostatic pressure treated follicles (33.3%) compared to control (20.2%);
group by TEM.
(p<0.05).
Conclusion: Hydrostatic pressure may play a critical role in oocyte
maturation. It can be a useful tool for the improvement of oocyte in vitro
maturation.
April 19-22, 2009 78

the receptor for melatonin, however the N-Acetylserotonin O-

P.020 – EMBRYO QUALITY IN POLYCYSTIC OVARY SYNDROME methyltransferase catalyzing the final reaction in melatonin biosynthesis is
actively expressed.
M. Balawi1,2, M. D. Cuquerilla2. 1Department of obstetrics and Gynecology.
General Hospital; 2Clinica Rubal, Gynecology and Reproduction; Ciuadad Conclusion: Modulators and growth factors generally described as
Real, Spain. effecteurs of nervous system are present in the oocyte. Some of them could
be considered like some members of the EGF network, as oocyte secreted
Introduction: polycystic ovary syndrome (PCOS) is the most common cause factors involved in cytoplasmic maturation. Receptors of neuromediators
of anovulatory infertility in women. Folliculogenesis in the PCOS ovary is are expressed: they are involved either in ion channel physiology and/or in
often disrupted, leading to suboptimal oocyte competence for fertilization. activation of cAMP dependant pathways. Based on their effect on early
This alteration in oocyte development is likely due to intrinsic molecular embryo development, in invertebrate, the implications for human assisted
defects in oocyte along with the state of androgen excess in PCOS patients. reproductive technology should not be overlooked.
Many therapeutic options are available to infertile couples with PCOS,
including controlled ovarian hyperstimulation and in vitro fertilization. The
aim of this study is to compare the embryo quality in women with PCOS and
women with normal ovaries undergoing in vitro fertilization (IVF/ICSI). P.023 – EFFECT OF SERUM PROTEIN SUBSTITUTE (SPS) ON THE MATURATION
Material and Methods: is a retrospective study conducted in Clinica Rubal OF HUMAN GV- OR MI-OOCYTES IN VITRO
during the period between 1st September 2007 and 30th December 2008. The Hwang-Yun Cho1, Jung-Lim Choi1, Seok-Yoon Lee1, Yong-Soo Heo1, Won-
number of embryos included in the study was 497 embryos, 249 coming Don Lee1, San-Huyn Yoon2, and Jin-Ho Lim2. 1Maria Fertility Hospital,
from 31 cycles in women with PCOS, and 248 coming from 50 cycles in 2
Fertility Research Center, Maria Medical Foundation, Seoul, Korea.
women with normal ovaries. To define patients with PCOS we used the
criteria of the consensus of Rotterdam 2003. We also used the criteria of Introduction In vitro maturation (IVM) of immature oocytes for infertile
ASEBIR (Spanish Association for the Biological study in Reproduction) to patients is an attractive treatment, because it can reduce side effects of
classify the embryo quality. ovarian stimulation with gonadotropins. However, there has been little
information about the suitable conditions for human IVM.
Results: The average age of women was 32.09 years for the PCOS group
and 33.8 years for the control group. Both groups were comparable. The Human follicular fluid (HFF) has been used in human IVF program as a
number of embryos, very good or good (A + B) in the PCOS group was protein source, but it is an undefined supplement. Because HFF has a risk of
higher, 177 (71%) compared with the control group 156 (62.9%) with a contamination and batch-to-batch variation, it should be replaced by
statistically significant P 0.02. However, the pregnancy rate was comparable synthetic serum substitute. This study was carried out to investigate
in both groups, 52% in the PCOS group vs. 51% in the control group, with whether serum protein substitute (SPS) is used as protein supplement for
no significant P. human oocyte maturation.
Conclusions: embryos coming from polycystic ovaries are not of inferior Materials and Methods Immature oocytes were obtained from 60 patients
quality compared with those from normal ovaries, and the pregnancy rate is underwent controlled ovarian hyperstimulation (from February to August
the same in women with or without polycystic ovaries undergoing IVF/ICSI 2008). A total of 280 immature oocytes, 183 of germinal vesicle (GV) and 97
of metaphase I (MI), were cultured in IVM medium (YS medium containing
7 IU/ml rFSH, 1 IU/ml rLH, and 10 ng/ml EGF) supplemented with either
P.021 – EXPRESSION OF NEUROMEDIATORS/EFFECTORS IN CYTOPLASMIC 40% (v/v) HFF or 40% (v/v) SPS (Sage/Cooper Surgical co.). Maturation
MATURATION OF HUMAN OOCYTES rates were compared between the two groups after 24 hrs.
Elisabetta Tosti, Yves Menezo, Moncef Benkhalifa. UNILABS, Laboratoire Results The maturation rate of SPS group (64.1%; 59/92) from GV oocytes
d'Eylau, Paris, France. was similar to HFF group (59.3%; 54/91). No difference was also the
Introduction: In the last years more and more attention has been paid to the maturation rate of MI oocytes between HFF (84.3%; 43/51) and SPS
effectors of oocyte maturation usually rather described as involved in the (89.1%; 41/46).
development of nervous system. Neuregulins have been shown to have Conclusion These results suggest that SPS should be use as a protein
diverse functions in the development of the nervous system. Brain-derived supplement for human oocyte maturation like HFF.
neurotrophic factor also known as BDNF is member of the "neurotrophin"
family of Nerve growth factors, found in the brain. For the neurotransmitters
we have looked at the mainly at GABA and serotonin, due to their P.024 – DEVELOPMENTAL POTENTIAL ON DAY 3 FAST-CLEAVING EMBRYOS
involvement in early embryonic development in invertebrates. AND PREGNANCY OUTCOME OF FAST-CLEAVING EMBRYOS
TRANSFER CYCLES
Material and Methods: According to the bioethical law, all the experiments
were performed on GV oocytes retrieved for ICSI, after conrolled ovarian Hye Won Choi1, Hee Jung Kang1, Mi Ra Shin1, Myo Kyung Kim1, Sun-Hee
stimulation. There is no transcription during the final stages of oocyte Lee1, Mi Kyoung Koong2, In Ok Song2, Chun Kyu Lim1; 1Laboratory of
maturation and the first embryonic divisions are under maternal control Reproductive Biology and Infertility, 2Department of Obstetrics and
before maternal to zygotic transition. So analysis of the oocyte gives an Gynecology, Cheil General Hospital & Women’s Healthcare Center,
interesting pictures of early embryonic cleavages.The protocol used for the Kwandong University College of Medicine, Seoul, Korea.
human oocytes has been already published. The analyses were performed, Introduction: It has been considered that 7~9 blastomeres are the optimal
on six pools of 10 oocytes, with microarrays, using the Affymetrix HG U-133 cell number of embryo for normal development on day 3. An association has
plus 2.0 chips. Amplification was performed via double IVT (in vitro been demonstrated between day 3 fast-cleaving embryos (≥ 10 cells) and
transcription, two cycles). blastocyst formation rate. We set out to determine the pregnancy potential
Results: Growth factors First of all, the neuregulins (NRG) 1 and 4 are of fast-cleaving embryos transferred on day 3.
expressed, but the intensity is 10 times higher for NRG1. NGFI-A binding Materials & Methods: Day 3 embryos were divided into three groups
protein 1 (EGR1 binding protein 1 and nerve growth factor receptor depending on their cell number: slow-cleaving embryos (4 ~ 6 cells),
(TNFRSF16) associated protein 1 are expressed in all the samples. normal-cleaving embryos (7 ~ 9 cells) and fast-cleaving embryos (≥ 10
Neurotransmitters: Serotonin 5-hydroxytryptamine (serotonin) receptor 7 cells). The blastocyst formation rate of each group was assessed in cycles
(adenylate cyclase-coupled) and 5-hydroxytryptamine (serotonin) receptor that the embryos were transferred on day 5 after oocytes retrieval. The
3, involved in depolarization of the membrane, are expressed. pregnancy outcome of each group was evaluated in cycles that embryos
were transferred on day 3 after oocytes retrieval. Only embryos of same kind
GABA GABA(A) receptor-associated protein-like 1, 2, and 3 are expressed. were transferred in each cycle and the pregnancy outcome was evaluated.
The solute carrier family 6 (neurotransmitter transporter, GABA), member
11 and the diazepam binding inhibitor (GABA receptor modulator, acyl- Results: Blastocyst formation rate of day 3 fast-cleaving embryos (30.0%)
Coenzyme A binding protein) are very significantly expressed . GABAA was significantly higher (P<0.01) than that of the slow-cleaving embryos
receptors are ligand-gated ion channels We did not find any expression of (16.9%). However, there was no significant difference between fast-cleaving
79 Geneva
embryos and normal-cleaving embryos (34.2%). A total of 50 embryos two groups (35.1% versus 41.7%; P=0.11). The cancellation rates were
(2.2±1.0) were transferred in 23 patients on day 3 and all of them were fast- also similar between two groups. (8.1% versus 6.3%; P=0.51).
cleaving embryos. The clinical pregnancy rate (12/23, 52.2%) and Conclusions: Young poor responders in the oc group showed a trend toward
implantation rate (19/50, 38%) were significantly higher (P<0.05) than them improved implantation and clinical pregnancy rates which did not achieve
(13.8% and 4.3%, respectively) of cycles that slow-cleaving embryos were statistical significance. Today both protocols accepted as viable options for
transferred. And the clinical pregnancy rate and implantation rate were poor responder patiens ≤37years.
higher than them (43.5% and 27.9%, respectively) of cycles that normal-
cleaving embryos were transferred. However, there was no significant
difference. Four patients have delivered six healthy babies and six P.026 – OUTCOME OF MILD STIMULATION CYCLES IN POOR RESPONDERS
pregnancies are ongoing. Two pregnancies have been aborted during first WITH SHORT-TERM APPLICATION OF GNRH ANTAGONIST AND LOW
trimester. DOSE GONADOTROPINS USING CLOMIPHENE CITRATE
Conclusions: According to our data, the developmental potential of day 3 Nur Dokuzeylul, MeteGurol U ur, Guvenc,Karlıkaya, Hale Karagozoglu, Aynur
fast-cleaving embryos is similar to normal-cleaving embryos. The day 3 Er ahin, M. Kavrut, Mustafa,Acet, Semra Kahraman; Istanbul Memorial
fast-cleaving embryo transfer cycles showed similar clinical pregnancy rate Hospital IVF and Genetics Center, Istanbul, Turkey.
and implantation rate to them of cycles that normal-cleaving embryos were
Introduction: The objective of this study is to evaluate the efficacy of
transferred. Our findings demonstrate that fast-cleaving embryos on day 3
minimal stimulation IVF with GnRH antagonist and rFSH or/and HMG in
have a comparable potential to normal-cleaving embryos in achieving
cycle using clomiphene citrate in young (37< years old ) and 38-42 years
pregnancies.
old poor responders.
Materials and Methods: We reviewed the medical records of all cycles
P.025 – LUTEAL ESTRADIOL/GNRH ANTAGONIST SUPPRESSION VERSUS ORAL performed in Istanbul Memorial Hospital ART and Genetics Department
CONTRACEPTIVE PRETREATMENT IN YOUNG POOR RESPONDERS from January 1, 2006 to, December 30, 2008. In a review of our clinical
Nur Dokuzeylul, MeteGurol Ugur, Guvenc Karlikaya, Hale Karagozoglu, Aynur data, we found 146 patients who were treated with clomiphene citrate (cc)
Ersahin, Mustafa Acet, Mehmet Karaca, Semra Kahraman. Istanbul and gonadotropins in young poor responders(Group A) and 150 patients in
Memorial Hospital IVF and Reproductive Genetics Unit, Istanbul, Turkey. 38-42 years old poor responders (Group B).Patients were included if they
were considered poor responders,as defined by one or more of the
Introduction: The purpose of this study is to compare cycle outcomes for following criteria (1)≤4 oocytes retrieved in previous stimulation;(2) basal
young poor responders(pr) treated with luteal estradiol patch (lep) FSH levels >10 mIU/mL (3)low estradiol(E2) level on the day of hCG
suppression versus an oral contraceptive (oc) pretreatment. administration (<800pg/mL) in previous stimulation. For both groups cc
Materials and Methods: We reviewed the medical records of all cycles was started on cycle day 3 and it has been continued for 5 days.On the 5th
performed in Istanbul Memorial Hospital IVF Unit from January 1, 2007 to, day of cc, the patients started taking gonadotropins and daily ganirelix
November 30, 2008. In a review of our clinical data, we found 37 patients acetate was started 0.25 mg subcutaneously when follicles reached 13-14
who were treated with lep and 79 patients who were treated with oc. mm in diameter. Ultrasound and estradiol measurements were used to
Patients were included if they were considered pr, as defined by one of the monitor follicular development. Oocytes were harvested 35-36 hours after
following criteria (1)≤4 oocytes retrieved in previous stimulation;(2) basal 10000IU hCG administration. Clinical pregnancy was defined as the
FSH levels >10 mIU/mL (3)low estradiol(E2) level on the day of hCG presence of a gestational sac on ultrasound.The primary end point was the
administration (<800pg/mL) in previous stimulation. For lep protocols, pregnancy rate. Data are compared by using student-t test and chi-square
patches containing 7.8mg estradiol/25cm2 was started on luteal day 21 and test.Results are presented as mean±standard error. P <0.05 was considered
it was replaced a new one two days later. Patients then remained on the last statistically significant.
E2 patch until the patch fell off or until day of hCG administration. On the 2nd Results: The mean age (33.08±3.30 vs. 40.10±1,36 years; P=0.0001) and
day of the transdermal E2 patch, the patients started taking antagonist daily the mean duration of infertility (8.00±4.85 vs. 11.13±7.10 years; P=0.0001)
ganirelix acetate 0.25 mg subcutaneously for 3 days. For oc protocol ,the pill were significantly different between two groups. Both groups were
containing gestodene 0.075mg and ethinyl estradiol 0.003 mg was started comparable with regards to mean number of previous trials (3.07±2.04 vs.
at 3. day of previous cycle and continued for 21 days. Ultrasound and 3.20±2.19 P=0.591), mean body mass index (24.78±4.64 vs. 26.32±4.44
estradiol measurements were used to monitor follicular development. kg/m²; P=0.91) and mean value of day 3 FSH levels(12.16±6.05 vs.
Oocytes were harvested by transvaginal ultrasound guided follicular 12.39±10.51 IU; P=0.213) respectively. Although there is no significant
puncture 35-36 hours after 10000IU hCG administration. Results are difference between two groups, the mean total amount of gonadotropins
presented as mean±standard error. P <0.05 was considered statistically required for controlled ovarian hyperstimulation in group B is higher than
significant. the group A (1434±999.83 vs 1426±1072.36 IU; P=0.37). There was also no
Results: Both groups were comparable with regards to mean age remarkable difference between the mean duration of stimulation period in
(30.84±0.48 versus 30.99±0.33years; P= 0.82), mean duration of infertility each group (4.47±2.32 vs 4.56±2.31; P=0.79). There were also no
(7.34±0.73 vs. 7.37±0.51 years; P=0.34), mean number of previous trials significant differences in the mean thickness of endometrium (8.21±2.13 vs
(2.30±0.20 vs. 2.14±0.11 P=0.37 ), mean body mass index (24.82±0.54 vs. 8.27±2.31 mm; P=0.313) and the mean serum estradiol (E2) levels, on the
24.79±0.47 kg/m²; P=0.47) and mean value of day 3 FSH levels(10.70±0.56 day of trigger of oocyte maturation between two groups (836±453.02 versus
vs. 10.37±0.40 IU; P=0.64) respectively. Although there is no significant 731±585.88 pg/ml; P=0.696). There were also no significant differences in
difference between two groups, the mean total amount of gonadotropins the mean number of retrieved oocytes (3.47±2.43 vs 2.79±2.60; P=0.622),
required for COH in lep group is higher than the oc group (4428.36±234.16 metaphase ΙΙ oocytes (2.76 ± 1.80 vs 2.38±1.97; P=0.32), the mean number
versus 4009.34±168.30 IU; P=0.27). There was also no remarkable of fertilized oocytes(2.33±1.53 vs 2.00±1.32; P=0.08). and the mean
difference between the mean duration of stimulation period in each group number of transferred embryos (1.91±0.94 vs 1.73±0.83; P=0.377). We
(9.32±0.28 versus 9.37±0.28; P=0.85). There were also no significant also found that there was no significant difference in the cancellation rates
differences in the mean thickness of endometrium (10.98±0.41 versus (21.4% vs 24.6%) and in the implantation rates between two groups (13.5%
10.68±0.18 mm; P=0.85) and the mean serum estradiol (E2) levels, on the vs 7.6%; P=0.095).The clinical pregnancy rates per ET(21.7% vs 10.6%;
day of trigger of oocyte maturation between two groups. (1173.29±101.47 P=0.03) and ongoing pregnancy per ET(16.5% versus 6.1%; P=0.02) are
versus 1230.63±65.036 pg/ml; P=0.11). There were also no significant significantly different between two groups.
differences in the mean number of retrieved oocytes (5.30±0.43 versus
5.63±0.28; P=0.61), M ΙΙ oocytes (4.63 ± 0.38 versus 4.67±0.24; P=0.94)
Conclusions: Young poor responders showed a trend toward improved
clinical and ongoing pregnancy rates by cc protocol.Thus it can be accepted
the mean number of PN (3.91±0.33 versus 4.03±0.21; P=0.62). and the as a viable option for young poor responder patients.
mean number of transferred embryos (2.32±0.14 versus 2.28±0.08;
P=0.62). Although the implantation rates and clinical pregnancy rates were
higher in the oc group,there were no significant difference in implantation
rates (22.2% versus 25.4%; P=0.11) and clinical pregnancy rates between
April 19-22, 2009 80

Materials and Methods: Normal Semen sample were collected by

P.027 – SOLUBLE FORMS OF FAS AND FAS LIGAND CONCENTRATIONS IN THE ejaculation, according to criteria of the World Health Organization (WHO), as
SEMINAL PLASMA OF SUBFERTILE MEN WITH OR WITHOUT well as spermatic morphology according to the strict Kruger criterion. Swim
VARICOCELE up technique was carried out to obtain 20 X 106 progressive motile sperms
with a good morphology. The motility speed of the preincubated
Yehia El-Garem1, Mohamed El-Sawi2, Adel El-Shafei1. 1 Department of spermatozoa introduced into the fertilization medium containing 0 mM or 5
Dermatology, Andrology & STD; 2Department of Clinical Pathology; Faculty mM and 10 mM caffeine were examined in capillary tube (25cm) as
of Medicine, Alexandria University, Alexandria, Egypt. cm/30minutes.
Introduction: The presence of a varicocele was often cited as the most Results: The results demonstrated that addition of caffeine with
common cause of reduced male fertility and decrease sperm quality. concentration 5mM to the fertilization medium show significant increase
The exact association between varicocele and male infertility is not well (P<0.05) in sperm speed motility from 7cm/30min to 11cm/30min. However
established. It is suggested, however, that varicocele is related to reduce higher concentration of caffeine (10mM) show no significant effect.
semen quality, decline in spermatogenesis. Conclusion: Caffeine stimulates human sperm hyperactivated motility. These
An altered apoptotic process has been found to be closely associated with results demonstrated that Caffeine enhanced several motion sperm
male infertility. The Fas system has been implicated as a key regulator of parameters and suggest a potential use of the Caffeine in infertile patients
germ cell apoptosis in the mammalian testis. with motility defects undergoing artificial insemination.
Up to 10% of sperm cells in the ejaculate of men with a varicocele were
apoptotic, as compared with 0.1% in fertile controls. That report concluded P.029 – COMPARISON OF TEST-YOLK BUFFER FREE SPERM
that varicocele induces apoptosis, which is initiated in the testicular tissue CRYOPRESERVATION MEDIA TO A TEST-YOLK BASED SPERM
and is then expressed in the semen. FREEZING MEDIUM
Soluble Fas (SFas) was found to inhibit Fas mediated apoptosis. The activity Samira Es-slami1, Susan Tarshala2, Richard Rawlins2, Rebecca Gilbert1.
of sFasL in inducing apoptosis has been extensively debated in the literature. 1
R&D Department, Irvine Scientific, Santa Ana, CA; 2Rush Center for
There is evidence both in favor of and opposing its apoptosis inducing Advanced Reproductive Care, Chicago, IL; USA.
activity. Recent reports suggest that human sFasL inhibits FasL-mediated
apoptosis, indicating that the shedding of FasL from the membrane to form Introduction: Sperm Maintenance Medium (SMM, Irvine Scientific) is an egg
SFasL is a mechanism for down-regulating of its killing activity. yolk free sperm cryopreservation medium containing glycerol and sucrose
as cryoprotectants and Human Serum Albumin (HSA). CryoSpermTM
Objective: The aim was to measure the soluble forms of Fas and Fas ligand (Medicult) is a protein free sperm cryopreservation medium containing
concentrations in seminal plasma of subfertile men with or without glycerol and raffinose as cryoprotectants. This study was conducted to
varicocele, to study the effect of varicocele on Fas pathway and compare the performance of these two Test-yolk buffer free media to test
spermatogenesis and correlate levels of sFas and sFasL with semen yolk buffer (TYB) containing medium: Sperm Freezing Medium-TYB (SFM-
parameters. TYB, Irvine Scientific).
Patients: Twenty subfertile male with varicocele (group A) and twenty Material and Methods: Semen samples were obtained from 25 patients. All
subfertile male without varicocele (group B) and ten fertile controls (group samples had normal concentrations and motility according to WHO
C). reference values (≥20 X 106 spermatozoa/ml and ≥50% Motile
Methods: All patients were subjected to semen analysis, hormonal assay spermatozoa). Routine semen analysis was performed under light
(FSH, testosterone) with chemiluminescence assay, scrotal duplex and microscopy to determine the motility and forward progressive motility (FP)
detection of seminal plasma levels of sFas and sFasL by double antibody of the samples. The semen from each patient was divided into equal
enzyme linked immunoassay. aliquots and cryopreserved in each of the 3 sperm cryopreservation media
according to the manufacturer’s instructions. After thawing, the % motility
Results: The levels of sFas ranged between 30-110 pg/ml in group A,
and % FP were determined for each sample, the % recovery post thaw was
between 90-202 pg/ml in group B, and between120-130 pg/ml in group C.
determined by comparing the pre-freeze and post thaw values. After removal
The levels of seminal plasma sFasL ranged between 0.8-2.6 pg/ml in group
of the cryoprotectants using a separation gradient, % motility and % FP post
A, between 0.9-1-6 pg/ml in group B and between 1.0-2.6 pg/ml in group C.
wash were determined. All assessments were made by the same observer.
Significant correlation between sFas levels and sperm count in the group A
Comparisons between groups were performed by using One Way ANOVA
(r=0.33, p=0.002)., There was positive significant correlation between sFas
followed by Student t test. A P value of <0.05 was considered statistically
levels and percent of rapid and slowly progressive sperms motility in group
significant. The results are presented as mean ± SEM.
A (p=0.03) and group B(p=0.01),while there was Positive significant
correlation between sFas levels and motile sperms in group A (r=0.44, Results: There was no significant difference in post thaw motility recovery
p=0.007) while there was insignificant correlation between sFas levels and and FP recovery between samples cryopreserved in SMM and SFM-TYB (%
percentage of motile sperms in the other 2 groups (p>0.05). There was motility recovery: 77±2.7 vs. 79±2.7; % FP recovery: 81±2.2 vs. 83±2.2).
negative significant correlation between sFasL levels and percent of motile Motility (%) and FP (%) post wash were similar as well (% motility: 87±1.1
sperms in group A (r=-0.3, p=0.05), group B (r=-0.3, p=0.03), group c (r=- vs. 87±1.1; % FP: 86±1.3 vs. 87±1.3). Samples cryopreserved in
0.4, p=0.05). No correlation between sFas, Fasl and sperm morphology and CryoSperm had significantly lower % motility recovery (62±3.6) when
with hormonal profile. compared to SMM and SFM-TYB. Percent FP recovery post thaw was lower
(75±2.5) than other media but did not reach significance. Post wash %
Conclusion: Apoptosis play an important role in pathophysiology of impaired
motility (78±3.2) and % FP (81±2.2) were both significantly lower when
spermatogenesis in subfertile patients with varicocele. SFas concentration in
compared to SFM-TYB and SMM.
the seminal plasma is a better marker for apoptotic changes than the sFasL.
The sFasL concentration in the seminal plasma implies the fine regulation of Conclusions: TYB based media have long been considered a first choice
SFasL in the function of the Fas system and consequently, of the apoptotic cryopreservation media for human sperm. In this study we showed that the
process in the human genital tract. performance of SMM, containing both glycerol and HSA without TYB, was
equivalent to TYB based medium SFM-TYB. The performance CryoSperm
was lower than SFM-TYB and SMM’s. The absence of Albumin in
P.028 – EFFECT OF CAFFEINE ON HUMAN SPERM MOTILITY IN VITRO CryoSperm could explain its lower cryoprotective ability.
Omar Elsraite, Mohamed Danfour, Bashir Awin, Mohamed Awin, Yousef
Awin, Ahmed Fakron, Mohamed Elmahaishi. Faculty of Medicine, 7th
October University - Misurata IVF Centre, Misurata, Misurata, Libya.
The effect of caffeine on the sperm motility was investigated after Incubation
30min In In Vitro Fertilization Medium.
81 Geneva
assisted reproductive treatments (ART) in Turkey.

P.030 – USE OF HUMAN-DERIVED FSH VERSUS RECOMBINANT FSH RESULTS Materials and Methods: A prospective study was carried out in a total of 105
IN MORE EMBRYOS SUITABLE FOR CRYOPRESERVATION men, of whom 43 with male factor infertility, 31 with female factor infertility
P. Fancsovits, GZs. Tóthné, Á. Murber, E. Hauzman, Z. Papp, J. Rigó Jr., J. and 31 with unexplained infertility diagnosis. The men answered
Urbancsek. First Department of Obstetrics and Gynaecology, Semmelweis questionnaires; the State Trait Anxiety Inventory(STAI), State Trait Anger
University, Faculty of Medicine, Budapest, Hungary. Scale(STAS) and the Beck Depression Inventory (BDI) during the treatment
cycle.
Introduction: The outcomes of IVF may change according to the type of
hormonal stimulation. However, there are very few studies analysing the Results: No significant differences between groups were found on measures
effects of different gonadotrophin preparations on oocyte and embryo of anxiety, depression or anger.
quality. Our prospective randomised study analyses the effect of highly Conclusion: In Turkey, male factor infertility as the cause of the couple’s
purified human-derived FSH (hFSH) and recombinant FSH (r-FSH) on oocyte infertility problem does not have an adverse effect on the psychological
and embryo quality in ICSI cycles. status of men undergoing ART. This study suggests that men’s
Materials and Methods: Seventy IVF cycles were randomly allocated to psychological adjustment to their own infertility diagnosis is not considered
human-derived gonadotrophin (Fostimon-HP®; IBSA, Lugano, Switzerland) to be psychologically ill.
or recombinant gonadotrophin (Gonal-F®; Serono, Rome, Italy) for ovarian
stimulation. Women with the following criteria were included in the study: P.032 – DOES CYTOKINE PROFILE IN SERUM AND FOLLICULAR FLUID DIFFER
infertility due to severe andrological factor, maternal age between 18 and 40 AFTER OVARIAN STIMULATION WITH GNRH AGONIST LONG OR
years, BMI 19–30 kg/m2, <3 previous completed cycles and basal FSH<10 ANTAGONIST PROTOCOL IN WOMEN UNDERGOING ASSISTED
IU/l within 3 months prior the study. Oocytes were fertilised by ICSI REPRODUCTIVE TREATMENT?
treatment. Oocyte morphology, fertilisation rate and embryo quality were
compared between the two groups. Assessment of oocyte morphology Banu Kumbak1, Oya Akcin1, Rukset Attar1, Gazi Yildirim1, Nihan Tecellioglu1,
included the most frequent oocyte dismorphisms, i.e., first polar body Serdar Oztezcan2, Cem Ficicioglu1;. 1ART Unit, Department of Obstetric and
abnormalities (fragmented, enlarged or immature), enlarged perivitelline Gynecology, Yeditepe University Hospital, Istanbul, Turkey 2Department of
space, considerable cytoplasmic vacuolisation or granularisation. Embryos Biochemistry, Yeditepe University Hospital, Istanbul, Turkey.
were graded according to their morphology observed at 42–48 hours and at Objective: To assess whether follicular fluid (FF) and serum (S) levels of
66–72 hours after ICSI. Morphological grading was based on the number cytokines in women undergoing assisted reproductive treatments (ART)
and size of blastomeres and the amount of fragmentation. In one r-FSH differ between GnRH agonist long and antagonist cycles.
cycle, where oocyte collection was performed, the male partner did not
Materials and methods: Eighty-five women who underwent ART either with
deliver a semen sample for fertilization and the oocytes of this cycle were
GnRH agonist long (n=34) or antagonist multiple-dose flexible protocol
not injected. Thus, this cycle was excluded from further analysis of oocyte
(n=51) were studied. FF samples were collected at the time of oocyte
and embryo morphology and pregnancy rate calculations. The Mann-
Whitney U-test and the χ2 test were used for statistical analysis.
retrieval and measured either by ezyme-linked immunosorbent assay
technique using commercially available kits [interleukin (IL)-1β, IL-6, IL-8,
Results: A total of 389 oocytes in 35 cycles were collected in the h-FSH IL-12, tumor necrosis factor (TNF)-α] or Griess method [nitric oxide (NO)].
group and 415 oocytes in 35 cycles were collected in the r-FSH group. Serum levels of those cytokines and NO were also assessed on the day of
Patients’ age, length of stimulation and number of gonadotrophin ampoules oocyte retrieval.
used for stimulation were similar in the two groups. The number of oocytes
Results: No significant differences were found in the FF concentrations of IL-
[11.1±3.9 vs. 11.9±4.1] and mature (MII) oocytes [9.9±4.1 vs. 10.6±4.3]
1β, IL-6, IL-8, IL-12, TNF-α and NO between the agonist long and
were not significantly different between the h-FSH and r-FSH group. The
antagonist cycles (p>0.05). The serum concentrations of those cytokines
frequency of morphologically abnormal oocytes (including abnormal first
were also similar in the two groups except NO (1.4±1.1 vs. 2.2±1.9 μM;
polar body [53.8% vs. 55.5%], enlarged perivitelline space [32.1% vs
p=0.038) and IL-6 (14.3±4.8 vs. 20.5±12.2 pg/ml; p=0.008) levels which
36.7%], cytoplasmic vacuolisation [7.1% vs. 4.1%] or granularisation
were found to be significantly lower in antagonist cycles.
[28.0% vs. 26.0%] were similar. The fertilisation rate was significantly
higher in the h-FSH than in r-FSH group [239/347; 68.9% vs. 217/362; Conclusion: It has been demonstrated that there is no significant difference
59.9%; P=0.01]. Morphological characteristics of embryos on day 2 and day in follicular microenvironment in terms of IL-1β, IL-6, IL-8, IL-12, TNF-α
3 were similar between the two groups. However, significantly more and NO levels between GnRH agonist long and antagonist protocols.
embryos were cryopreserved in the h-FSH group [91/239, 38.0% vs. However, serum IL-6 and NO levels which were found to be significantly
62/217, 28.6%; P=0.03]. Positive βhCG-tests [16/35, 45.7% vs. 14/34, lower in women from antagonist group might be of clinical value which
41.2%], clinical pregnancy rates [13/35, 37.1% vs. 11/34, 32.4%] and should be evaluated with future investigations.
implantation rates [17/109; 15.6% vs. 20/111; 18.0%] were similar between
the two groups.
P.033 – MEIOTIC AND REDOX CONSEQUENCES OF GLYCOLYSIS INHIBITION IN
Conclusions: Stimulation with recombinant gonadotrophin for ICSI BOVINE CUMULUS-OOCYTE COMPLEXES DURING IN VITRO
treatment seems to result in a similar number of oocytes and similar oocyte MATURATION
morphology as stimulation with highly purified human-derived
Cynthia Gutnisky1, Gabriel Dalvit1, Jeremy Thompson2, Pablo Cetica1.
gonadotrophin. Oocytes obtained with h-FSH stimulation showed higher 1
Institute of Research and Technology in Animal Reproduction (INITRA),
fertilisability, whereas embryo morphology, pregnancy and implantation
Area of Biochemistry, School of Veterinary Sciences, University of Buenos
rates did not differ between the groups. However, patients might benefit
Aires, Argentina; 2The Robinson Institute, School of Paediatrics and
from h-FSH stimulation because of a higher proportion of embryos suitable
Reproductive Health, The University of Adelaide, Australia.
for cryopreservation.
Glucose is necessary for cumulus-oocyte complex in vitro maturation.
Glucose catabolism by glycolysis within cumulus cells produces ATP and
P.031 – PSYCHOLOGICAL INFLUENCE OF MALE FACTOR AS THE CAUSE OF substrates that can be subsequently oxidized by the oocyte. In other somatic
THE COUPLE’S INFERTILITY ON MEN WHO ARE UNDERGOING cells, the glycolytic pathway is negatively regulated by ATP by inhibition of
ASSISTED REPRODUCTIVE TREATMENTS: A PRELIMINARY STUDY IN phosphofructokinase. Sodium fluoride (NaF) can also inactivate the pathway
A TURKISH POPULATION by inhibiting the glycolytic enzyme, enolase. The intermediary metabolism of
Banu Kumbak Aygun, Irem Atak, Rukset Attar, Gazi Yildirim, Narter glucose also produces NADH, which, besides being used for metabolism,
Yesildaglar, Ates Karateke, Cem Ficicioglu. Department of Obstetrics and also helps regulate the oocyte REDOX potential. This work studied the effect
Gynecology, Yeditepe University Hospital, Istanbul, Turkey. of the inhibition of glycolysis during in vitro maturation on glucose uptake
and lactate production of COCs and the nuclear maturation rate of the
Objective: To investigate the psychological influence of male factor infertility
oocyte. Because of the results obtained, a further aim was to determine the
as the cause of the infertility problem on men in couples undergoing
effect of inhibition of glycolysis on oocyte REDOX state and rate of meiotic
April 19-22, 2009 82

maturation. COCs were recovered by aspiration of antral follicles (2-5 mm in the mean number of obtained oocytes, fertilized, fertilization rate was
diameter) and only oocytes completely surrounded by a compact and (12.7±5.3, [n= 3038]); 7.3±2.8; [n=1755]; 57.8%). A total of 1210
multilayered cumulus oophorus were used. Glucose uptake and lactate pronucleate oocytes embryos were frozen and 864 were thawed. However,
production from spent media was measured by spectrophotometric assays only 592 were survived (survival rate 68.5%). 22 pregnancies were achieved
following culture of individual COCs in 20 µl drops of maturation media. (9.2%; p=0.068). In both groups The pronucleate stage embryos survival
Maturation was carried out in medium 199 supplemented with 5 % fetal rate after freeze thawing procedure was 67.1% and the pregnancy rate was
bovine serum, FSH, LH and gentamycin (control) and ATP (1, 10, 20 and 40 13.6%.
mM) or NaF (2, 3, 4, 5 mM). After 22 h of maturation, COCs were removed Conclusion: Despite the indications and techniques of becoming pronucleus
from culture to evaluate the rate of meiotic completion to the metaphase II stage embryos were different, the survival and pregnancy rate was similar
chromosome configuration. In order to study the redox potential of the and added more than 10% pregnancy rate for ART program.
oocyte, groups of 50 COCs were cultured in maturation media described
above (control) supplemented with 10 mM ATP or 3 mM NaF. The redox
level of the oocyte and the maturation stage was measured at 4 different P.035 – URINARY HMG FOLLOWING RECOMBINANT FSH FOR CONTROLLED
time points (0 – 9 – 15 – 22 h). Oocytes were stained with the fluorescent OVARIAN HYPERSTIMULATION YIELDS BETTER CLINICAL OUTCOMES
probes, Redox Sensor Red CC-1 and MitoTracker Green and observed by THAN RECOMBINANT FSH ALONE IN GNRH ANTAGONIST IVF /ICSI
laser scanning confocal microscopy. The images were analysed using CYCLES
Photoshop 9.0 software. To assess the maturation stage, oocytes were
Atsushi Haruki, Tomoko Inoue, Fujio,Migishima, Keijiro Ito, Hirotsugu Oku,
stained with 10 µg/ml Hoechst and evaluated under fluorescence
Yoshiharu Morimoto. IVF Namba Clinic, Osaka City, Osaka province, Japan.
microscopy. COCs matured in the presence of ATP and NaF showed a dose
dependant inhibition in glucose uptake and lactate production (p<0.05), and Introduction: Recombinant FSH has an important role for controlled ovarian
in their progression to metaphase II (p<0.05). Oocytes showed changes in hyperstimulation in IVF/ICSI cycles and GnRH antagonists have been
their REDOX level throughout maturation (p<0.05). However, these changes introduced in many IVF centers. But it has not yet become evident whether
were not observed either with ATP or with NaF (p<0.05), and an inhibition in recombinant FSH alone or urinary hMG for controlled ovarian
the migration of mitochondria was observed in the presence of these hyperstimulation is more effective in GnRH antagonist IVF/ICSI cycles. This
compounds (p<0,05). High positive correlations between the REDOX level study was aimed at comparing the two preparations for controlled ovarian
and mitochondrial activity were observed (r>0.82; p<0.05). Both, ATP and hyperstimulation in patients with a GnRH antagonist protocol.
NaF blocked nuclear maturation at the germinal vesicle stage (p<0.05). Material and Methods: In the retrospective study, we analyzed a total of 154
These results demonstrate that glycolytic pathway activity is essential for patients following a GnRH antagonist IVF/ICSI cycle from June 2006 until
bovine oocyte in vitro maturation. The oocyte REDOX levels vary during the June 2008. The exclusion criteria were age > 40 years and WHO Group I
maturation process and they are related to mitochondrial activity. The anovulatory women. Oral contraceptives were administered to all women
inhibition of nuclear maturation by ATP and NaF may be linked to changes in during the prior period of treatment cycles. The patients were divided into
the REDOX potential and the inhibition of the migration of mitochondria. two groups by means of gonadotropic preparations for ovarian stimulation.
Ovarian stimulation was initiated with recombinant FSH from day 3 of the
cycle for both groups. For one group, recombinant FSH was given
P.034 – PREGNANCY RATE OF FROZEN-THAWED PRONUCLEUS STAGES
continuously until the day of inducing ovulation (rFSH group). For the other
EMBRYOS FROM IVF AND ICSI
group, recombinant FSH was followed by urinary hMG at least from the day
M.E. Hammadeh, M. Deryal, A.S. Amer, C. Fischer-Hammadeh. Department 5 of cycles until the day of inducing ovulation (hMG group). Cetrorelix in the
of Obstetrics and Gynaecology, University of Saarland, Homburg/Saar, dose of 0.25mg was started to be administered daily when the leading
Germany. follicle reached 14 mm. When at least two follicles reached a mean diameter
Introduction: The purpose of this study was to determine and compare the of >17mm, 5000IU of HCG was given and oocytes were retrieved 36-38
survival and pregnancy rate of frozen-thawed pronucleate stage embryos of hours after HCG injection. A good-quality embryo was defined by their
IVF and ICSI patients. morphologic features and appropriate cleavage rate. A good morphology of
blastocyst was defined as having a well-expanded blastocoele cavity, a well-
Material and Methods: 431 women underwent either IVF (n=191) or ICSI defined inner cell mass and a single layer of trophectoderm cells
(n=240). All women received GnRHa for down regulation and rec FSH surrounding the cavity. After conception, we followed the patients until 8th
(Gonal-F) for controlled ovarian hyperstimulation. After 18-24 hours of weeks of pregnancy. Clinical pregnancy was defined as presence of a
oocyte insemination (IVF) or sperm injection (ICSI) the pronucleate stage gestational sac.
embryos were randomly assigned to be cryopreserved or to continue
development for potential embryo transfer (≤3Embryos) according to the Results: In both groups, no significant difference was observed in the
female age (cut-off point =35 year). Pronucleate stage embryos were frozen background factors of the patients as an age, the past number of IVF/ICSI
in phosphate–buffer saline (PBS) medium containing 1, 2 propanediol treatment cycles, basal FSH level. There was no significant difference in the
(PROH), sucrose and 20% patients’ serum in three steps (0.5 mol PROH, serum estradiol and progesterone levels measured on the day of hCG
1.0 mol PROH, 1.5 mol PROH plus 0.1 mol sucrose) at room temperature injection, a number of mature oocytes and the fertilization rate. A
for 10 min. each. The freezing process was initiated at 24 hour after significantly longer duration of stimulation (9.6 versus 8.4 days) and thus a
insemination or oocytes injection. The mini straws were cooled slowly from significantly increased requirement for gonadotropic dose was present in
22 °C until -7°C (2 °C /min.), after holding them at this degree for 10 min., the hMG group. A rate of good morphology blastocyst was similar among
they were cooled further until -30°C at a rate of -0.3 °C/min. followed by the two groups, but a rate of good-quality embryos in the rFSH group was
cooling until -170 °C/min. at 20°C /min. They are then plunged into liquid higher than that in the hMG group (31.5% versus 15.0%; P=0.06). Clinical
nitrogen for storage. Seeding was done at -7°C. After thawing of pronucleate pregnancy rate per embryo transfer (20.0% versus 39.4%; P=0.06) and the
stage embryos by removing the straw from liquid nitrogen into water bath rate of early pregnancy loss (28% versus 8.3%) for the rFSH and hMG
30 °C. The cryoprotectants were removed in four steps dilution using PBS groups respectively differed in favor of the hMG groups.
with PROH and sucrose. The pronuclear stage oocytes are inspected for Conclusions: A quality of the embryos in the rFSH group was higher than in
survival before being incubated for 24 hours. Embryo transfer was the hMG group. However, a lower ongoing pregnancy rate was seen in the
performed a 24 h culture. Up to three pronucleate embryos were transferred rFSH group, most probably due to a worsened environment of implantation
only if cleavage takes place. including endometrial receptivity.
Results: The mean number of obtained oocytes, fertilized, fertilization rate of
IVF patients was (12.9±5.7; [n=2458]; 8.9±4.2, [n=1701], 69.2%). However,
1210 pronucleate stage embryos were frozen, 850 were thawed. Besides,
558 were survived the freeze-thaw process (65.6%. survival rate). This
survived Pronucleate stage embryos were cultured until the first or second
division was completed (24 hours), Embryo transfer was performed at 24
hours after thawing. 34 women became pregnant (17.8%). In ICSI program
83 Geneva
Conclusion: This study indicated that pregnancy rate per cycle was 11.8%.
P.036 – LEUKOCYTOSPERMIA AND ITS CORRELATION WITH SPERM IUI success rate decreased when the woman age was more than 35 years
PARAMETERS IN PATIENTS REFERRED TO KASHAN INFERTILITY old or sperm count < 10 × 10 6 . 10 million sperm must be Inseminated
CENTER DURING 2007-2008 When the normal morphology was less than 50%.
Hassan Hassani Bafrani1, Razieh Afrugh2, Fatemeh Fruzanfard3. 1Department

of Anatomy, 2School of Medicine, 3Department of Obestetric & Gynecology, P.038 – OBSTETRIC OUTCOMES OF PREGNANCIES FROM IN-VITRO
School of Medicine, Kashan University of Medical Sciences, Kashan, Iran. MATURATION OF OOCYTES TREATMENT
Introduction: The reactive oxygen species (ROS) derived from abnormal H.M. Tuong, V.T.N. Lan, D.Q. Vinh, P.H. Tuan. IVF Van Hanh, Ho Chi Minh,
sperm or from WBC or both of them. ROS in semen is responsible for the Vietnam.
lipid peroxidation, sperm motility defect, and decreases the sperm Introduction: In-vitro maturation (IVM) of oocytes has been developed
fertilization ability. The aim of this study was to determine sperm quality recently and has become more popular at IVF centers worldwide. There have
(count, motility, morphology) in infertile men with or without been few data on the pregnancy and obstetric outcomes of babies born from
luckocytospermia and abnormal patterns of ROS production in. this technique. This study is to report the obstetric outcomes of the first
Materials and Methods: We retrospectively reviewed semen analysis of 110 cohort of babies following IVM treatment in Vietnam.
infertile men referred to Infertility Center during 2007-2008. Material and methods: This was an observational study conducted on
Semen was centrifuged at 1200 ×g for 5 min to separate seminal plasma. women conceived and then gave births from IVM treatment due to the
The aliquots were stored at −80 °C until analyzed. indication of PCOS at IVF Van Hanh, Ho Chi Minh City, Vietnam from
Seminal plasma and serum were resuspended in phosphate buffer saline for September 2007 to September 2008. Pregnant women were examined for
MDA levels in seminal plasma by spectophotometric assays. weight gain; complications occurred during pregnancy such as pre-
Results: After semen analysis in 110 patients, the semen samples were put eclampsia and diabetes mellitus. Ultrasonography was used to investigate
in luckocytospermia group (n=30), and nonluckocytospermia group (n=35). number of fetus, nuchal translucency, and 3-D morphology of the fetus. The
In nonluckocytospermia leukocyte number was <0.25 × 106 babies born from IVM treatment were recorded with regards to mode of
delivery, gestation at birth, number of babies, birth weight, gender, apgar
Sperm motility had significantly different between leukocytospermia and score, and complications at birth such as respiratory distress syndrome,
nonluckocytospermy patients, and also in healthy men (P<0 .05 and P< intracranial hemorrhage, and major birth defects.
0.001 respectively).
Results: There were 31 women conceived and gave births, amongst them,
ROS level also had significantly difference between leukocytospermia and there were 20 singleton and 11 twin-pregnancies. The mean female age was
healthy men, and between nonluckocytospermy patients with healthy men 29.8 ± 2.8 years. The mean weight gain was 15.0 ± 4.7 kg. No case of pre-
(P<0 .001 and P< 0.0001 respectively). eclampsia and diabetes mellitus was recorded. There was no case of
Conclusions: With respect to correlation of leukocytospermia with sperm abnormal nuchal translucency and 3-D morphological ultrasonography. A
motility, and important role of sperm motility in fertilization, we can total of 42 babies were born from the IVM treatment, in which, there were
conclude that detection and treatment of leukocytospermia, after excluding 18 male (42.9%) and 24 female (57.1%). Two cases had vaginal
genitourinary infection, may have positive influence on fertility and spontaneous delivery, others had cesarean sections. There were 6 preterm
pregnancy rates in infertile couples. In respect of relationship between births; the mean gestation at birth was 37.6 ± 1.9 weeks (30 – 40 weeks).
sperm motility and leukocytospermia, and beneficial effect of motility, The mean birth weight was 2602 ± 480 g (1700 – 3450 g). Two babies had
leukocytospermia can affect fertility capability patners. However, application apgar score at 1 minute of 6, and 40 babies had score of >/= 7. No severe
of antioxidant in leukocytospermia treatment, but final effect of the complications and major congenital defects were recorded at births.
antioxidant is still controversial. Conclusion: Pregnancies from IVM treatment has good obstetric outcomes.
Data from the first cohort of babies born from IVM program in Vietnam are
reassuring for women undergoing IVM treatment.
P.037 – INVESTIGATING OF SPERM PARAMETERS AND PREGNANCY
OUTCOME AFTER INTRAUTERINE INSEMINATION
Hassan Hassani Bafrani1, Maasoomeh Abedzadeh2, Fatemeh Fruzanfard3. P.039 – THE IMPACT OF SERUM AND FOLLICULAR FLUID ANTI-MULLERIAN
1
Department of Anatomy, 2School of Medicine, 3Department of Obestetric & HORMONE LEVELS ON IN VITRO FERTILIZATION OUTCOMES IN
Gynecology, School of Medicine, Kashan University of Medical Sciences, NORMAL AND POLYCYSTIC OVARY SYNDROME WOMEN
Kashan, Iran. S. Arabzadeh1, G. Hossei1, B. Rashidi2, M. Agha Hosseini3. 1School of
Introduction: Infertility prevalence is about 10-15 percent. IUI is one of the Biology, University College of Science, University of Tehran; 2Imam
treatment methods in infertility. The IUI success rate is Influenced by Khomeini Hospital, Vali-e-Asr Reproductive Health Research Center, School
different factors. Different studies indicated that Induction of ovulation and of Medicine, Tehran University of Medical Science, 3Shariati University
IUI increased the chance of pregnancy in the infertile couple special with Hospital IVF center, School of medicine, Tehran University of Medical
specially male factor infertility. The aim of this study was to investigate the Science; Tehran, Iran.
success rate of IUI with related factor in the kashan fertility and sterility Introduction: Anti-mulleian hormone (AMH) is a biomarker that predicts the
center. number of antral follicles and is involved in follicle arrest for women with
Material and Methods: 84 infertile couple that underwent 102 IUI cycles with polycystic ovary syndrome (PCOS). This study investigates the relationship
washed husband semen were included in this study. All patients’ charts between serum or follicular fluid (FF) AMH concentrations and In Vitro
were reviewed for age, IUI number, semen characteristics and pregnancy Fertilization (IVF) outcomes in normal and PCOS women.
rate. Material and Methods: Women with PCOS, (20-47) years, (n=33), and
Findings: Total pregnancy rate were 11.8 percent per cycle and 14.3% per normal ovulatory control, (21-42) years, (n=56), body mass index 21-33
infertile couple. The women age mean in pregnant group was 25.8 ± 26 kg/m2 in PCOS vs. 19-35 kg/m2 in normal controls were recruited from Vali-
years old and in non pregnant group was 29 ± 5.3 years old. The t-student e-Asr Reproductive Health Research center and Shariati University Hospital
test in both groups were significant. The majority of pregnancies were IVF center during this retrospective study. On the cycle day 3, serum
Achieved in women less than 35 years old (13.3%). Induction ovulation samples and on the day of oocyte retrieval FF from more than one
protocol in 66/7% women was HMG. All of pregnancies were achieved in preovulatory follicle (16-20 mm in diameter) was collected for AMH
couples with first IUI cycle. measurements by using Elisa method. Association between AMH levels and
oocyte number, oocyte maturation, fertilization rate, implantation rate,
Post wash semen parameter including : Sperm count ≥ 10 × 10 6 , motility ≥
embryo grade, biochemical and clinical pregnancy were assessed with one-
50%, normal morphology ≤50% were observed in 91.7%, 100% and
sample Kolmogorov-smirnov, spearman, Mann Whitney and binary logistic
83.3% of pregnancy rate respectively but difference statistically were not
regression statistical tests.
significant in two groups.
April 19-22, 2009 84

Results: Median (range) serum AMH level was markedly increased in the achieved successful IVF outcome compared to other E2/MII ratio
PCOS group [15.06 (0.10-50.70) vs. 3.38 (0.42-9.91) in normal controls; categories.Consequently, this retrospective study implies that the decision of
P<0.0001]. Similarly, median (range) FF AMH level was higher in PCOS appropriate HCG administration is not so critical in GnRH-agonist protocol,
group (8.21 (0.39-127.59) vs. 2.13 (0.62-13.69) in normal controls; but is very critical in GnRH-antagonist protocol to obtain best IVF-ET
P<0.05). In both groups, serum AMH levels showed a significant positive outcome.
correlation with oocyte number (r = 0.40; P<0.01 vs. r = 0.49; P<0.01 in
PCOS) and oocyte maturation (r=0.46; P<0.0001 vs. r = 0.59; P<0.0001 in
PCOS). Embryo grade remained irrespective of AMH concentrations in P.042 – COMPARISON OF CUMULATIVE PREGNANCY RATES IN IVF-ET CYCLES
serum or in FF in both normal control and PCOS groups. There was an BETWEEN CULTURE OF ALL FERTILIZED EGGS AND SPLIT ZYGOTES
inverse relation between fertilization rate and serum AMH levels (r = -0.29; AFTER FREEZING AT SIBLING PRONUCLEAR STAGES IN THE PATIENTS
P<0.05); a positive relation between FF AMH levels and implantation (r = HAVING SMALL NUMBER OF ZYGOTES
0.32, P<0.05) and biochemical pregnancy (P<0.05) rates in normal control Myo Kyung Kim1, Hee Jung Kang1, Dong-Wook Park1, Seung Bum Hong1, Mi
group but not in PCOS group. However, no significant correlation was Ra Shin1, Inn Soo Kang2, Jin Young Kim2, Chun Kyu Lim1. 1Laboratory of
observed between serum or FF AMH levels and clinical pregnancy in both Reproductive Biology and Infertility; 2Department of Obstetrics and
groups. Gynecology, Cheil General Hospital & Women’s Healthcare Center;
Conclusion: Concentration of AMH in serum, but not in FF, may constitute a Kwandong University College of Medicine, Seoul, Korea.
useful marker of oocyte number and oocyte maturation in PCOS and normal Introduction: It is well known that cryopreservation of supernumerary
control group. While, AMH levels in FF could be a predictive factor of embryos could increase the cumulative pregnancy rates. However, there is
implantation and biochemical pregnancy rates in normal control but not in no consensus regarding cryopreservation at the PN stages to improve
PCOS women. pregnancy outcomes in IVF cycles with a small number of zygotes. This
study was carried out to see whether cryopreservation of split PN zygotes
could increase the cumulative pregnancy rates in the patients having eight
P.040 – SERUM ESTRADIOL VALUE AND ESTRADIOL / METAPHASEIIOOCYTE 2PN-zygotes.
RATIO ON THE OUTCOME OF IN VITRO FERTILIZATION WITH
CONTROLLED OVARIAN HYPERSTIMULATION Materials and Methods: This retrospective study analyzed IVF or ICSI cycles
carried out between Jan. 2003 to Dec. 2007. We selected 138 embryo
Mamoru Ida1, Fumie Nagata1, Kengo Sugihara1, Yoshiharu Nakaoka1, Aisaku transfer (ET) cycles that were estimated eight zygotes (2PN) after 20-22
Fukuda1, Yoshiharu Morimoto2. 1IVF Osaka Clinic; 2IVF Namba Clinic; Osaka, hours insemination/ICSI and less than nine fertilized zygotes (included 1PN
Japan. or delay developed embryos). Total ET cycles were divided into two groups:
Introduction: The aim of the present study was to evaluate the peak estradiol group I (n=86); total fertilized embryos were cultured to transfer on day 3
(E2) value on the day of human chorionic gonadotropin(HCG) administration without PN stage freezing. Group II (n=52); after some sibling zygotes were
and the ratio of E2 per number of metaphaseIIoocytes retrieved (E2/MII frozen at the PN stages, the others were cultured to day 3 until to transfer.
ratio) during the controlled ovarian hyperstimulation (COH) with Clinical pregnancy outcomes were compared with between two groups in
gonadotropin-releasing hormon(GnRH)-agonist or GnRH-antagonist, on the fresh ET cycles and cumulative ongoing pregnancy rates were estimated
outcome of in vitro fertilization-intracytoplasmic sperm injection (IVF-ICSI) after subsequent frozen-thawed ET.
cycles. Results: There were no significant differences in female mean age, number
Materials and Methods: Retrospective analysis was performed on the of retrieved oocytes and total fertilized embryos between two groups.
patients who completed IVF-ICSI cycles from January 2006 to July 2008. Number of cultured embryos was significantly lower in group II (5.2±0.5)
The elimination of bias in this selection was achieved by excluding either than in group I (8.4 ± 0.7) (P<0.01). Also, number of transferred embryos
poor responders or high responders: women who achieved E2 levels was significantly lower in group II (3.3 ± 0.6) compared with group I (3.6 ±
<500pg/ml or >5000pg/ml on the day of HCG. Other exclusion criteria were 0.6) (p<0.01). The number of beta-hCG positives and delivery rates (51.2 vs
frozen-thawed embryo transfer cycles. Two COH protocols were used: 537 46.2 % and 47.7 vs 44.2%) after fresh ET were slightly higher in group I
patients were treated with the midluteal long GnRH-agonist protocol than in group II. The differences were not statistically significant. However,
(agonist group) and 162 patients with flexible multidose GnRH-antagonist the cumulative ongoing pregnancy rates after frozen-thawed ET cycles were
protocol (antagonist group).The selection of types of analog was largely slightly higher in group II (51.9%) than in group I (47.7%).
dependent on clinical characteristics of the patients. Each group was divided Conclusions: Cryopreservation of sibling zygotes could slightly increase
into four subgroups according to their peak E2 value on the day of HCG: cumulative pregnancy rates in the patients having less than ten fertilized
≤1000pg/ml; 1001-2000pg/ml; 2001-3000pg/ml; >3000pg/ml.Additionally eggs compared with all zygotes cultured group. This IVF-ET policy could
each group was divided into another four subgroups according to their offer higher chances to cumulative pregnancies and the patients could have
E2/MII ratio: ≤100pg/ml;101-200pg/ml;201-300pg/ml; >300pg/ml. Number benefit on cost effectiveness.
of MIIoocytes retrieved, number of embryos transferred and pregnancy
rates were assessed .
Results: Pregnancy rate per transfer cycles was significantly higher in P.043 – COMPARISON OF CLINICAL OUTCOMES ACCORDING TO DAYS OF
agonist group (45.4%) than in antagonist group (33.3%) EMBRYO TRANSFER IN ART CYCLES
(p,<0.01).Pregnancy rates in E2 value of ≤1000pg/ml, 2001-3000pg/ml, and Sun-Hee Lee1, Hee Jung Kang1, Dong-Wook Park1, Hye Won Choi1, Seung
>3000pg/ml were comparable between agonist and antagonist groups, while Bum Hong1, Hae Ok Kim2, Chan Woo Park2, Chun Kyu Lim1. 1Laboratory of
pregnancy rate in E2 value of 1001-2000pg/ml was significantly higher in Reproductive Biology and Infertility, 2Department of Obstetrics and
agonist group than in antagonist group(p<0.05).In both agonist and Gynecology, Cheil General Hospital & Women’s Healthcare Center,
antagonist groups, there was no difference in pregnancy rates between Kwandong University College of Medicine, Seoul, Korea.
different E2 subgroups.Pregnancy rates in E2/ MII ratio of 101-200pg/ml
Introduction: The majority of embryo transfers (ETs) to date have been
and 201-300pg/ml were comparable between agonist and antagonist
performed on day-3 to reduce potential risks for developmental arrests of in
groups, while pregnancy rates in E2/MII ratio of ≤100 pg/ml and >300pg/ml
vitro cultured embryos before ETs. Development of sequential media has
were significantly higher in agonist group (p<0.05 for both).Furthermore,
significantly improved culture conditions that could allow for blastocyst
there was no difference in pregnancy rates among the different E2/MII ratio
transfer on day-5. While day-5 ETs provide higher clinical pregnancy
subgroups in agonist group. However, higher pregnancy rate was observed
outcomes with reduced risks of multiple pregnancies, they still have
in E2/MII ratio of 101-200 pg/ml comparing to those in ≤100 pg/ml or
potential risks of developmental arrests of IVF embryos. The aim of this
>300pg/ml in antagonist group (p<0.05 for both).
study was to evaluate clinical outcomes of day-4 ETs and compare the
Conclusions: The peak E2 value on the day of HCG cannot predict clinical efficacy of day-4 ETs with other days of ETs in our center.
outcome either in agonist group or antagonist group. While E2/MII ratio
Material and Methods: From January 2006 to August 2007, total 1124
cannot predict clinical outcome in agonist group, only the patients
cycles were analyzed – 897 day-3, 121 day-4, and 106 day-5 ETs. The cycles
undergoing GnRH-antagonist protocol with E2/MII ratio of 101-200 pg/ml
85 Geneva
with number of retrieved oocytes < 5, female age >37 and/or any genetic agonists in combination with recombinant follicle stimulating hormone in –
factors were excluded. The rates of matured oocyte, fertilization, good 30%.
embryo, and clinical pregnancy were examined among three groups. Chi Results: After prompt analysis the following results were registered:
square test and ANOVA were used for statistic analysis. P values < 0.05 pregnancy rate after IVF was 30,9% and after ICSI 31,8%; delivery rate after
were considered significant. IVF and ICSI were 26,24% and 27,9% accordingly; mean number of
Results: There were no significant differences among three groups with embryos transferred in each cycle were 2,3 and 2,4 after IVF and ICSI (we
respect to mean age of female and rates of matured oocyte, fertilization and would like to mention the fact that after September 2007 maximal number of
good embryo. Clinical pregnancy rate of day-4 ETs (55.4%) was significantly embryos transferred were equal to 2). As to deal with embryologic stage we
higher than that of day-3 ETs (41.5%) (p=0.0051). There was no significant had transfer of embryo only on day 3 in 21,7% of cases, on day 5 in –
difference in clinical pregnancy rates between day-4 and day-5 ETs. 39,8%; and on day 3 and 5 in – 38,4%. In all our embryo transfer
Conclusions: Day-4 ETs had clinical pregnancy rates as high as day-5 ETs procedures we used COOK catheters. After embryo transfer procedure for
and it is significantly higher than that of day-3 ETs. Day-4 ETs may provide a luteal support were used utrogestane and crynone (8%) in 79% and 21% of
better option to achieve a high clinical pregnancy outcome with reduced cases accordingly. From all attempts first attempt was effective in 64% of
risks of multiple pregnancies and developmental arrests of embryos. women, second – in 17,6%; third – in 9,8%; effectiveness of four and more
attempts was less then 2% per cycle. We also tried to examine effectiveness
of attempts in accordance to age of the patients. The data were following: in
P.044 – OUTCOME AFTER HYSTEROSCOPIC CORRECTION OF UTERINE women younger then 27 years pregnancy rate was equal to 36,9%; in group
ANOMALIES of 28-35 years – 30,5%; in 36-40 year group – 26,4%; women older then
41 years – 15,8%. When analyzing the number of embryos transferred per
Kay Jayakrishnan, RamAnupama. KJK Hospital, Trivandrum, Kerala, India.
cycle we found out that after transfer of three embryos pregnancy rate was
Introduction: Mullerian anomalies are the frequent congenital malformation 34,7%; delivery rate – 29,5%; number of multiple pregnancies – 15%. In
of the genital tract. It is associated with various types of reproductive failure, contrary after transfer of 2 embryos we had the further results: pregnancy
including recurrent miscarriage,late abortion and preterm delivery . rate was 30,9%; delivery rate – 26,2%; number of multiple pregnancies –
Methods: 123 women with mullerian anomalies were detected during a eight 9%. From all started cycles 317 (26,24%) cases ended with delivery of
year period from January 2001 to December 2008 in a prospective cohort babies. Among all these were diagnosed 27 (8,5%) multiple pregnancies.
study. The anomalies were detected by pelvic sonography including 3 D Conclusion: Differential approach to IVF programme optimizes and leads to
scan, diagnostic hysteroscopy, hysterosalpingography and MRI scan. Of this increase of IVF results.
52 patients (55% ) presented with primary infertility and 42 patients (44 %
) with secondary infertility.In this study we have excluded male factor
infertility, unmarried patients and other causes of infertility. These patients P.046 – THE EFFECTS OF SPERMPREPARATION ON THE INITIATION OF
were subjected to laparohysteroscopy .Complete septum in 59 CAPACITATION OF HUMAN MALE GAMETES
patients(62.7%) and partial septum in 8 (8.5%)cases. 13 patients had Gudrun Keck, Nadja Gneist, Evelyn,Gouma, Katrin Hanseroth, Birgit
unicornuate uterus with rudimentary horn, 6 patients had arcuate uterus,3 Leuchten, Berit Thieme, Ina,Trinkaus, Wolfgang Distler. Universitatsklinikum
had bicornuate uterus,3 had mullerian agenesis and 2 presented with Dresden IVF-Labor, Dresden, Sachsen, Germany.
bicornis unicollis. Surgical intervention was in the form of septal resection
in 67 (71.2%) of cases, lateral metroplasty in 6 (6.3%) cases of unicornuate Introduction: Glycodelin (GD) is a progesterone-regulated lipocalin protein
uterus, rudimentary horn excision in 3 (3.1%), right rudimentary horn with of the reproductive axis with diverse actions in cell recognition and cell
hematometra excision in 1case and vaginoplasty in a single case. differentiation. It is a 28 to 30-kD glycoprotein synthesized in various
Conception rate following surgical intervention, effort of cervical encerclage glands, notably in the male and in the female reproductive organs.
on pregnancy, surgery to conception interval, mode of conception and route Glycodelin appears in different glycoforms on sperm surface and in seminal
of delivery were followed up in our study group. plasma (GD-S), in amniotic fluid (GD-A), in follicular fluid (GD-F) and in
cumulus cells (GD-C).
Results: 33conceptions (35.1%) occurred in our study group. 90.9% of our
conceptions within 1 year after surgery and rest within 4 years after surgery. GD-S bound to the surface of human spermatozoa is known to inhibit the
17 patients (51.5%) conceived spontaneously, 9 (27.2%) following capacitation of sperms, which is a prerequisite for fertilization. Thus, it
controlled ovarian stimulation,6 (18.1%) patients following controlled needs to be removed from the surface to enable the capacitation of the
ovarian stimulation combined with IUI and 1 case following in vitro sperms. In in-vivo-conception GD-S is naturally removed during the passage
fertilization. We have done cervical encerclage in 8 (11.9%) cases of septum through the cervical mucus. In contrast; up today there is nothing known
resection. All of them delivered at term ( 100 % live birth rate . These about the fate of GD-S during in vitro fertilization. Therefore the aim of this
results were statistically significant.About 19(57.5%) delivered by caesarean study was to investigate whether or not GD-S is removed during the semen
section and 7( 21.2%) by vaginal route. We had a follow up percentage of preparation procedure prior in vitro fertilization.
68.08%. Materials and Methods: Ejaculates were assessed of men attending our IVF
Conclusions: Endoscopic surgery restores normal uterine cavity, reduces program. The study was approved by the Ethics Committee of the University
morbidity and post operative adhesions and thereby improves the Hospital Dresden. All patients gave signed informed consent. Only ejaculates
conception rates. from patients with normozoospermia were included in the study (nEJ = 32).
Semen preparation was carried out via swim up procedure (SUP). The
amount of bound GD-S on sperm surface was investigated on each 200
P.045 – DO WE HAVE RESERVE TO RAISE EFFECTIVENESS OF IVF? sperms prior and each 200 sperms after SUP per ejaculate. GD-S was
visualized by immunocytochemistry staining with a monoclonal antibody
Leonid Kuzmichev, Elena Kalinina, Veronika Smolnikova, Marina Shakhova,
Mf8 against the glycodelin protein core. The stainings were imaged and the
Khatuna Surmava, Liya Kazaryan. Moscow, Russia.
GD-binding on spermatozoa was semi-quantified by scoring (nSP =12 800).
Introduction: In our centre from May 2006 to may 2007 were performed The amounts of bound GD both on sperm surface in ejaculate and in sperm
1208 IVF cycles. suspensions after SUP were correlated (SPSS 12.0).
Materials and Methods: Among these female factor of infertility was Results: With the method established in our workgroup it was possible to
diagnosed in 35% of cases, male – in 34% and both factors were present in detect and to semiquantify bound GD-S in both; the fresh ejaculates, as well
21% of cycles. One attempt of IVF was performed in 27,4% of patients; two as on the surface of sperms SUP. The results of the scoring showed a
attempts in - 30,2%; three attempts in - 28,8%; fore and more attempts - in decreased amount of bound GD-S after SUP in all investigated ejaculates.
13,3% of all cases. The protocols of ovarian stimulation we have used were The decrease was statistically significant (p< 0.05). The findings indicate
the following: antagonists in combination with human menopausal that semen preparation via SUP significantly reduces the amount of bound
gonadotropins in 23,1% cases, antagonists in combination with GD-S on the surface of sperms, enabling capacitation of sperms even in
recombinant follicle stimulating hormone in – 24,7%; agonists in vitro.
combination with human menopausal gonadotropins – in 25,8% cases;
April 19-22, 2009 86

Conclusion: With these results we could prove for the very first time that an surgically retrieved sperm with no significant difference (p = 0.944). The
important prerequisite for initiation of capacitation of human sperms, the pregnancy rate is 34.8% using fresh surgically retrieved sperm and 39.4%
removal of GD-S, can be achieved in- vitro by the swimming up semen using frozen-thawed one.
separation method. Conclusions: The fertilization and pregnancy rates after ICSI using fresh and
frozen-thawed surgically retrieved sperm were comparable. Thus, it shows
P.048 – IMMUNOCOMPETENT CELLS OF PLACENTA FROM WOMEN OF HIGH the chance of achieving a clinical pregnancy following ICSI was not
FERTILITY AGE IN CASE OF PATHOLOGICAL PREGNANCY correlated to the status of the sperm whether it is fresh or frozen.
Cryopreservation of surgically retrieved sperm did not alter pregnancy
Natalia Linkova, Artem Kostylev, Irina Kostyuchek, Viktoria Polyakova, outcome. Recovery of surgically retrieved sperm for cryopreservation from
Natalia Palchenko. Ott Institute of Obstetrics and Gynecology, Saint- the husband is strongly recommended before ovarian stimulation as
Petersburg, Russian Federation. instances have been found of failure to obtain sperm on day of oocyte
Introduction: Women older than 30 years (high fertility age) frequently have collection, hence, the oocytes will not be wasted. Also, it is certainly a valid
difficulties in pregnancy such as: threat of an abortion, spontaneous option in these group of patients as ICSI may be performed later or even in
abortions, gestosis, uterine inertia, fetal intrauterine hypoxia. Thus, another center using the frozen-thawed surgically retrieved sperm without
pregnancy and labors in this age are often links with changes in the system jeopardizing the success rate.
“mother- placenta-fetus” and can be the cause of obstetric and perinatal
pathology. The pathological processes are accompanied with changes of
immune status, which, in turn, can be the important indicator of those P.050 – MATURITY OF OOCYTE MAY IMPACT OUTCOME OF IN VITRO
violations. MATURATION DURING CONTROLLED OVARIAN HYPERSTIMULATION
FOR PREVENTING SEVERE OVARIAN HYPERSTIMULATION SYNDROME
The aim was to study various immunocompetent cells in placentas of
Soo Jin Chae1, Chang Young Hur2, Kyung Sil Lim3, Jin Ho Lim4. Maria
killers), Т-killers, Т-supressors, B-lymphocytes and plasma cells in a
women older than 30 years. Our task was detection of NК-cells (natural
Fertility Hospital, Seoul, Korea.
placentas of women in chosen age category. Introduction: In vitro maturation (IVM) during controlled ovarian
Materials and Methods: 10 placental biopsies from women with a various hyperstimulation (COH) is effective method for prevention of ovarian
pathology (6 had gestosis of light and middle degree and 4 - had other hyperstimulation (OHSS). We investigated the factors which may influence
pregnancy pathology). The average women’s age was 33,7 years (from 30 clinical pregnancy of IVM during COH for preventing OHSS.
(plasma cells), СD20 (B-lymphocytes), CD8 (Т-killers and Т-supressors),

till 38 years). Immune cells were identified by according markers: CD40 Materials and Methods: We included the women who had undergone OHSS
in the previous COH cycles or had susceptibility for OHSS between January,
CD4 (Т- supressors) and CD16 (natural killer cell). Slides with tissue 2006 and December, 2007 by review of medical record in Maria fertility
specimens were photographed at augmentation 400x. Quantitative analysis hospital. Women with only first autologous cycle of IVM during COH were
of immunohistochemical expression was done by using computer image enrolled. After at least 5 days’ stimulation with gonadotropin, if the diameter
analisator «Video-Test, Morphology-5». Relative square (S, %) and intensity of leading follicle 12-14mm with more than 20 growing follicles, 10,000IU
of a staining (I, relative units) were estimated. hCG was injected. After 36-38hr, oocytes were aspirated using 19 gauge
Results: NK-cells were founded in placenta (SCD16=3.36±1.13 %, needle. Immature oocytes filtered using mesh were cultured with YS
ICD16=0.42±0.10). The large value of intensity and low values of square can medium with 1 IU/ml FSH, 10 IU/ml hCG and 10ng/ml recombinant human
be connected with high localization’s degree of NK-cells. Though NК-cells epidermal growth factor. After denudation with hyaluronidase, ICSI was
compound only 10-15 % from a total number of lymphocytes, they realized performed. Zygotes were co-cultured with cumulus cells in 10μl YS medium
such important function as destruction of target-cells without direct contact, with 10% human follicular fluid and were transferred. We compared the
clinical pregnancy group and non-pregnant group according to the
immune response which realized by Т- and B-lymphocytes. Small amount of
with the help of perforine protein, and, accordingly, without development of
developmental and clinical variables.
cells, expressing CD8 (SCD8=1.9±1.6%, ICD8=0.22±0.17) were also Results: A total of 110 patients received IVM during COH cycles (103 for
indicates on absence of Т-supressors. Positive response at CD8 is cause of

detected in placental tissue. We suppose that absence of CD4 in placenta cleavage stage embryo and 7 for blastocyst). Overall ongoing pregnancy
presence only Т-killers or low differentiated T-lymphocytes.

rate was 34.5% (38/110). Minimal to moderate OHSS was developed in
4.5% (5/110) and severe OHSS was not shown. Clinical pregnancy group
(n=44, 40%) showed similar clinical variables such as age (mean±SD;
with immune responses by NK-cells and only insignificant part realize by Т-
Conclusion: Pathological pregnancy for women of high fertility age connects
32.0±2.9 years vs. 32.1±3.2 years), FSH (5.7±1.8 mIU/ml vs. 4.5±1.4
killers. mIU/ml), number of previous IVF cycle (1.0±1.5 vs. 0.7±1.2), etiology of
infertility, total dose of gonadotropin (16.2±6.7 ampoules vs. 18.1±7.5
ampoules), duration of stimulation (6.9±1.2 days vs. 7.2±1.8 days),
P.049 – COMPARISON OF FERTILIZATION AND PREGNANCY RATES AFTER ICSI endometrial thickness at transfer (9.4±1.5mm vs. 9.9±1.5 mm), and number
WITH FRESH AND FROZEN-THAWED SURGICALLY RETRIEVED SPERM of embryos transferred [cleavage; 4.6±0.9 (n=40) vs. 4.7±0.9 (n=63),
blastocyst; 2.3±0.5 (n=4) vs. 2.3±0.6 (n=3)] compared with non-pregnancy
Shaw Ni Amy Lee, M.N. Lim, C.F. To, S.L. Yu. Department of Obstetrics &
group (n=66). Number of MII oocytes to those of total oocytes retrieved
Gynaecology, CARE, Singapore General Hospital, Singapore.
ratio is significantly higher in clinical pregnancy group [479/968 (49.5%)]
Aim: This retrospective study aimed at comparing the results of than non-pregnancy group [408/980 (41.6%), P<0.001]. Day 1 fertilization
intracytoplasmic sperm injection with fresh and frozen-thawed sperm rate was significantly different between clinical pregnancy group [365/479
obtained after microsurgical epididymal sperm aspiration (MESA) and (76.2%)] and non-pregnancy group [337/408 (82.6%), P=0.019]. Clinically
testicular sperm extraction (TESE) in azoospermic patients. pregnant women showed no significant difference in total fertilization rate
Materials and Methods: A total of 56 fresh cycles of ICSI treatment were [603/798 (75.5%) vs. 646/823 (78.5%)], day 2 fertilization rate [208/282
carried out using fresh and frozen-thawed surgically retrieved sperm (73.8%) vs. 264/355 (74.4%)], and day 3 fertilization rate [30/37 (81.1%)
between February 1994 to December 2008, 23 using fresh retrieved sperm vs. 47/70 (67.1%)] compared with non-pregnant women.
and 33 using frozen-thawed retrieved sperm. ICSI was performed using Conclusions: Women with more mature oocytes may show higher
these sperm on the wives’ oocytes after undergoing controlled ovarian pregnancy rate of IVM during COH than those with less mature oocytes
hyperstimulation and oocyte recovery with ultrasound guidance. A despite lower initial fertilization rate. Maturity of oocytes may be a predictor
maximum of 3 embryos were selected for transfer on either Day 2 or Day 3 for clinical pregnancy of IVM during COH for preventing severe OHSS.
after oocyte recovery. Retrospectively, the fertilization and pregnancy rates
were compared. Results were analyzed statistically using chi-square test.
Results: No significant difference was observed in the fertilization rate using
fresh and frozen-thawed surgically retrieved sperm (60.3% versus 55.5%);
p = 0.270. The pregnancy rate is similar using both fresh and frozen-thawed
87 Geneva
higher (P<0.05) compared with corresponding rate in the single step

P.051 – USING HIGH MAGNIFICATION MICROSCOPY TO EVALUATE FREZZING procedure.
EFFECT ON SPERM HEAD INTEGRITY Conclusion: The results of present study indicated that germinal vesicle
Gemma López, Rafael Lafuente, Sergi Rovira, Olga Cairó, Mario Brassesco. oocytes vitrified stepwise procedure had positive effect on survival,
CIRH. Clínica Corachan. Barcelona, Spain. maturation, fertilization cleavage and blastocysts rate than single step
procedure.
Introduction: Evidence in long IVF series suggests that sperm morphology,
according to a strict criteria, predicts an important information about fertility Key words: Germinal Vesicle Oocyte, Vitrification, mouse, Ethylene glycol,
rates. The aim of this study is to evaluate the effect of freezing on sperm blastocyst.
head integrity after capacitation by gradient method, analyzing the presence
of nuclear vacuoles. P.053 – PROSPECTIVE RANDOMIZED STUDY TO COMPARE THE EFFICACY OF
Materials & Methods: Prospective study that analyses samples from RECOMBINANT HUMAN CHORIONIC GONADOTROPIN (R-HCG) VERSUS
accepted donors after a preliminary study in our clinic. We have analysed URINARY HCG (U-HCG) IN FINAL FOLLICULAR MATURATION OF 365
the sample before 1 hour post-ejaculation and the same sample once PATIENTS UNDERGOING ASSISTED REPRODUCTIVE TECHNOLOGIES
thawed, after be prepared with density gradients and frozen. The analysis (ART)
was made in real time through inverted microscope Leica AM 6000 with Zeng Yong1, Hu Xiaodong1, Mo Meilan1, Song Cheng1, Zhang Wei M. Meilan1
100x immersion objective and 1.6x multiplier (8000x). A total of 100 Ye Xingshen2, Diego Ezcurra3. 1Shenzhen Zhongshan Urology Hospital,
spermatozoids were counted in every sample, and divided into the following China; 2 Merck Serono China; 3Merck Serono SA, Geneva, Switzerland.
categories:
Objective:To compare oocyte and embryological outcomes of 365 IVF cycles
Group I: without nuclear vacuoles triggered with r-hCG versus u-hCG
Group II: 1 or 2 small vacuoles
Group III: 1 large vacuole Design: Prospective randomized study
Group IV: many small vacuoles Materials/Method: Three hundred and sixty five infertile patients from the
Group V: 1 large vacuole and other vacuoles Center of Assisted Reproductive Techniques, Shenzhen Zhongshan Urology
Results: Average of all samples analysed Hospital - China, eligible for IVF/ICSI treatment, participated in this study. All
patients were treated with long pituitary down-regulation with the injection
Fresh sample: 12.7% of group I; 25.5 % of group II; 30.3% of group III; of 1.25 mg of GnRHa in the mid-luteal phase of the preceding cycle. Five
17.16% of group IV; 14.33% of group V. days after menses, ultrasound scanning was performed to confirm the
Prepared and Frozen samples: 6% of group I; 25.3% of group II; 27% of achievement of down-regulation (absence of an ovarian cyst and
group III; 23% of group IV and 18.7% of group V. endometrial thickness of less than 6mm). Controlled ovarian
Comparing between the fresh samples and frozen ones, we observe a hyperstimulation (COH) was initiated with a daily dose of 150-375 IU of
reduction in the number of spermatozoa without vacuoles, and a light recombinant human FSH (r-hFSH) or human menopausal gonadotrophin
increasing of spermatozoa with large vacuoles and with numerous small (hMG). The ovarian response was monitored by ultrasound scanning.
ones, in those samples analysed after thawing. Urinary LH level was tested when the leading follicle was ≥ 14 mm. Cycles
were cancelled if follicles remained with a diameter of ≤10 mm after 14 days
Conclusions: By means of sperm magnification we have observed that after of stimulation. Patients with more than two follicles ≥18 mm, were
capacitation and freezing there is an increasing of spermatozoa presenting randomized to receive 250 μg of recombinant human chorionic
altered morphology. We continue studying further and trying to evaluate its gonadotrophin (r-hCG) by subcutaneous injection or 10,000 IU of urinary
repercussion on fertility. human chorionic gonadotrophin (u-hCG) by intramuscular injection.
Transvaginal ultrasound-guided oocyte retrieval was scheduled 36 hr after
hCG injection and all visible follicles were aspirated. Fertilization was
P.052 – IN VITRO MATURATION, FERTILIZATION AND EMBRYO
performed by IVF or ICSI (patients with poor sperm quality). Embryos with
DEVELOPMENTAL CAPACITY OF MOUSE GERMINAL VESICLE OOCYTES
≥6 blastomeres and ≤20% fragments were defined as good quality.
FOLLOWING STEPWISE CRYOPRESERVATION
Continuous variables were expressed as mean ± SD and were compared
Reza Mahmoudi1, Mina Dehghani2, Hamdollah Delaviz1. 1Department of between the two groups using Student’s t-test. Categorical variables were
Anatomy and Embryology, Medicine Faculty, Yasuj University of Medical compared using Chi-square test. Differences were considered significant
Science, Yasuj; 2Dept. of Surgery, Namazi Hospital of Shiraz University, when the p value <0.05.
Shiraz; Iran.
Results: The characteristics of IVF cycles utilizing r-hCG and u-hCG were
Backround: Cryobiology is a very important tool in reproductive biology. compared (Table 1). The results expressed as mean ± SD or percentages,
This procedure can benefit the cancer patient wishing to preserve fertility as appropriate, are summarized below.
before initiation of any destructive chemotherapy or radiation therapy. It is a
Table 1
substitute for embryo cryopreservation and thereby avoids associated
ethical issues. Oocyte cryopreservation technology can lead to the Variable uhCG 10.000 IU Ovidrel 250 mcg P value
establishment of “oocytes banks” and provides solutions to ovarian failure Number of patients 181 184
patients. Age (years) 32.2 ± 5.6 32.0 ± 5.8 0.739
Years of infertility 4.6 ± 3.2 4.8 ± 3.5 0.606
Objective: The aim of this study was to evaluate viability, fertilization and
Mean gonadotropin utilized 43.2 ± 14.5 41.6 ± 15.9 0.328
subsequent developmental to blastocyts of mouse germinal vesicle oocytes
(75 IU vials)
after single and stepwise vitrification procedure.
Days on treatment 11.9 ± 2.5 12.0 ± 2.1 0.840
Material and Methode: Oocytes were obtained from 3- 4 week old female Number of oocytes retrieved 13.3 ± 6.0 12.9 ± 6.5 0.521
mice 48hr after i.p. injection of 7.5 IU pregnant mare serum gonadotropin MII ratio (MII/oocytes retrieved) 94.45% 95.07% 0.480
(PMSG). Collected oocytes before vitrification were exposed to Fertilization ratio IVF 84.37% 82.68% 0.246
cryoprotectant, which was composed of 30% (v/v) ethylene glycol, 18% (oocytes fertilized/oocytes inseminated)
(w/v) Ficoll-70, and 0.3 M sucrose, either by single step or in a step-wise Cleavage rate 97.61% 97.96% 0.583
way. After vitrification and storage in liquid nitrogen, the oocytes were Embryo excellence ratio IVF 41.45% 44.70% 0.297
thawed and washed two times in medium TCM199 and then subjected to in (good quality embryos/all embryos)
vitro maturation, fertilization and culture for blastocysts. Embryo excellence ratio ICSI 38.02% 44.78% 0.010
Results: The oocytes survival rates after vitrifying-warming, maturation rate,
the capacity of fertilization and embryonic development to blastocyst were Conclusions: The number of oocytes retrieved and the percentage of those
examined in vitro. The oocytes surviving, maturing to MII, fertilization, that were mature (MII ratio) were not significantly different when triggering
cleavage and blastocyst rate in the step-wise exposure were significantly final follicular maturation with recombinant or urinary hCG. Cleavage rate
April 19-22, 2009 88

was not different for both treatment groups and embryo excellence rate Materials and Methods: The testis of mouse minced into small pieces
(good quality embryos/total embryos produced), in ICSI patients, was mechanically and enzymatic digestion was performed to separate the cells
significantly lower with urinary hCG than the comparator. The rest of the from seminiferus tubules. Lectin-coated dishes used for sertoli cell isolation
variables analyzed were not significantly different between the two treatment from spermatogonial stem cells. With the formation of a confluent layer of
groups. The results of this study suggest that r-hCG is effective for sertoli cells, spermatogonial stem cells transferred on this feeder layer. Also
achieving final follicular maturation and the production of good quality mitomycin C-treated STO fibroblast cell line was used. In control group,
embryos in patients undergoing ART. spermatogonial stem cell cultured on a feeder free culture dishes. The
number and diameter of mouse spermatogonial stem colonies was assessed
after 3, 7 and 10 days of culture by an inverted microscope.
P.054 – WHEN SHOULD WE FREEZE ALL ZYGOTES TO PREVENT OHSS IN IVF
CYCLES WITH OVARIAN STIMULATION, E2 VALUE OF HIGHER THAN Results: The results of one-way ANOVA and Tukey post hoc test showed
5000PG/ML OR 3500PG/ML? significant differences between the mean number and diameter of the
colonies between sertoli, STO and control groups in 7th and 10th days
Yukiko Miyaki1, Asuka Ohtani1, Kengo Sugihara1, Mamoru Ida1, Yoshiharu (p<0.05), however there wasn’t significant difference between these three
Nakaoka1, Aisaku Fukuda1, Yoshiharu Morimoto2. 1IVF Osaka Clinic; 2IVF groups in 3th day (p>0.05).
Namba Clinic; Osaka, Japan.
Conclusions: The present study demonstrated that Sertoli cell may have
Introduction:Controlled ovarian hyperstimulation (COH) increases the more positive influence on the process of in vitro colonization.
number of oocytes retrieved and is making it possible to select better quality
embryos for embryo transfer (ET). On the other hand, ovarian
hyperstimulation syndrome (OHSS) is one of the most serious side effects. P.056 – EVALUATION OF SERUM ANTI-MULLERIAN HORMONE(AMH) LEVEL IN
Cryopreservation of all embryos is one of the alternatives to avoid THE IVF
aggravation of OHSS and also implantation failure due to extremely high Miyuki Nagaki, Kaori Goto, Yoko Kumasako, Eiko Otsu, Takafumi
estrogen. Our present indication for cryopreservation of all zygotes is E2 Utsunomiya. St-Luke Clinic, Oita, Japan.
value of higher than 5000pg/ml on the day of HCG administration. In the
present study, we investigated the occurrence of OHSS and pregnancy rate Introduction: The age of infertility patient when start the treatment is
in fresh cycles among three different ranges of E2:3500-4000pg/ml, 4000- increasing older because an age at marriage is going to up in Japan. As a
4500pg/ml and 4500-5000pg/ml to reevaluate the appropriate E2 value for new marker to examine the ovarian reseve, Anti-Mullerian hormone(AMH)
cryopreservation of all zygotes to prevent the occurrence of OHSS. was reported. The AMH level in in-vitro fertilization was investigated.
Material and Methods: Sixty eight fresh ET cycles (65 cases) with E2 values Materials and Methods: A total of 172 cycles of patients who scheduled in-
between 3500 and 5000pg/ml on the day of HCG administration from vitro fertilization from January 2008 to July 2008 were devoted to our study.
January, 2006 through December, 2007 were analyzed retrospectively. All The blood sampling for the AMH measurement was performed at the ovary
cases were stimulated by gonadotropins with GnRH analog. COH started on stimulation was started (on the fifth day of menstrual cycle).
day 3 of menstrual cycles and ovarian follicles were monitored by
AMH levels were measured duplicately and caliculated the mean AMH level.
transvaginal ultrasonography from day 7. When more than two follicles
reached 18 mm in diameters, 5000 unit of HCG was administered. Oocytes AMH levels in the serum were assayed by a commercially available enzyme-
were retrieved 36 hours after HCG administration. Cleavage or blastocyt linked immno sorbent assay(ELISA) kit, AMH used leader 680 model
stage embryos were transferred and occurrence and intensity of OHSS were microplates (BIO-RAD company), and absorbance was measured at 450nm.
followed for 4 weeks after transfer. All cases were divided into 3 groups by Results: The mean age of 172 cycles of patients who measured the AMH
the values of E2 on the day of HCG (Group A: 3500 < E2 <4000, Group B: level was 36.4±4.4 years old.
4000 < E2 <4500 and Group C: 4500 < E2 <5000). The occurrence of OHSS
As the patient’s age became higher, the mean AMH level were decreased
with it’s intensity and pregnancy rate in each group were compared.
significantly (The mean AMH level of patients who are less than 34 years
Results: The mean age of the patients and the number of embryos old : 2.68ng/ml , 35 - 39 years old : 1.83ng/ml, more than 40 years old :
transferred among these three groups were similar. There were no 1.03ng/ml).
significant differences in pregnancy rates among Group A (57.5%: 23/40),
When a high number of follicles were recognized at the time of the oocyte
Group B (59.1%: 13/22) and Group C (50.0%: 3/6). Occurrences of OHSS
pick up, AMH level were increased(Less than 3 follicle : AMH0.45ng/ml, 4-6
were comparable among Group A (20.0%: 8/40), Group B (27.3%: 6/22)
follicle : AMH1.27ng/ml, 7-9 follicle : AMH2.13ng/ml, 10-15 follicle :
and Group C (33.3%: 2/6) and there were no significant differences of their
AMH3.08ng/ml, More than 16 follicle: AMH4.1ng/ml).
intensity.
In 159 stimulation cycles, the AMH level of the no-oocyte group(AMH :
Conclusions: There were no differences of pregnancy rates and also
0.33ng/ml) of 24 cycles was compared with the normal group (AMH :
occurrences of OHSS among the three E2 values from 3500pg/ml to
2.10ng/ml) of 135 cycles. The AMH level tended to be lower in the no-
5000pg/ml on the day of HCG administration. The present study reaffirmed
oocyte group than the normal group.
that the present our criteria, E2 value higher than 5000pg/ml, for freezing
total zygotes to prevent OHSS is appropriate. However, we should follow The AMH level was compared in 76 cycles of embryo transfer (ET) group
very carefully the IVF patients with peak E2 value of 3500pg/ml or higher on with 27 cycles of ET cancelled group(not including all 2PN stage freeze
the day of HCG in stimulation cycles. because of avoidance from OHSS). The AMH level was significantly lower in
ET cancelled group(AMH: 0.33 ng/ml) than ET group (AMH: 1.73ng/ml).
Embryo growth on the third day after oocyte pick up showed better growth
P.055 – THE EVALUATION OF THE TIME EFFECT OF CO-CULTURE SYSTEM ON
when AMH was high level, there was not a significant difference(The
SPERMATOGONIAL STEM CELL COLONIZATION
percentage of good embryos of the AMH level 1ng/ml group: 75.0%, the
M. Mohamadi1, M. Movahedin1, M. Koruji2. 1Department of Anatomy, Tarbiat AMH level 1 - 2ng/ml group: 85.7%, the AMH level more than 2ng/ml group:
Modares University. 2 Basic sciences, University of Social Welfare and 95%).
Rehabilition Sciences; Tehran, Iran.
The pregnancy rate per stimulation cycle with an AMH level less than 1ng/ml
Introduction: Spermatogonial stem cells are unique population of cells with group was 9.8%, In contrast, the pregnancy rate with an AMH level more
self-renewing potential, and are the only stem cells in the body that transmit than 1ng/ml group was 25%. It showed a significant difference.
genetic information to the next generation and can be cultured for extended
Conclusion: In IVF treatment, the AMH level on the fifth day of a menstrual
periods in the presence of feeder cells such as sertoli and STO cells.
cycle(before HMG start), will be able to predict the IVF outcome.
Objective: This study aimed to compare the effect of co-culture with sertoli
and STO cells on the number and diameter of mouse spermatogonial stem
cell colonies to improve an in vitro culture system capable of supporting
spermatogonial stem cell colonization.
89 Geneva
and was decided considering female age and cause of infertility, thus not all
P.057 – 3D POWER DOPPLER MEASUREMENT OF PERIFOLLICULAR the surviving oocytes were inseminated. Cook Cleavage Medium was used
VASCULARIZATION IN NORMAL AND POLYCYSTIC OVARIES DURING IN for embryo culture. Embryo transfer (ET) was done on day 2-3 after
VITRO FERTILIZATION insemination. ßhCG determination was performed 15 days after ICSI and
clinical pregnancy was assessed by transvaginal ultrasound 3 weeks later.
A. Nazzaro1, A. Salerno1, M.D. Limongelli 1, G. Carlomagno2.
1
Physiopathology of Human Reproduction Unit, AORN “G. Rummo” Results: A total number of 524 MII oocytes from 58 cycles were frozen and
Hospital, Benevento; 2Department of Obstetrics and Gynecology, University in 25 cycles 193 were thawed. Oocyte survival rate was 41% (79/193). The
of Rome “La Sapienza”, Roma; Italy. mean number of microinjected oocytes was 3.6 ± 1.4 and fertilization rate
was 69.1 %. A mean of 2.3 ± 1.2 embryos were transferred. Embryonic
Objective: Ovarian angiogenesis and subsequent intrafollicular O2 content is development rate was 93.6 % from which 34.1 % corresponded to embryos
a critical point in determining oocyte competence and IVF outcome. We of good morphologic quality and appropriate cleavage. No significant
wanted to evaluate the differences, if any, in ovarian perifollicular blood flow differences on embryo quality was found between fresh cycle and frozen-
(PFBF) in normal and polycystic ovaries (PCOS) during IVF by using three- thawed cycles (p<0.2). Nineteen cycles reached ET. The 6 failed cycles were
dimensional power Doppler (3D pD). due to: no surviving oocytes (2), no fertilization (2) and arrested embryos at
Materials: 80 unexplained infertile women and 74 women with ultrasound pronuclear stage (2). The mean age of patients who had ET was of 33.4 ±
evidence of PCOS were enrolled. Controlled ovarian hyperstimulation (COH) 4.4 years. Implantation, clinical pregnancy and live birth rates per ET were
was induced by long or short protocol + rFSH( -follitropin). A GE 14.0 %, 31.6 % and 21.1 %, respectively. One ongoing pregnancy and four
730Voluson machine with a transvaginal volume transducer was used. liveborns were achieved. No multiple pregnancies occurred. The cumulative
3DpD was used to assess total follicle number per ovary (FNPO), ovarian pregnancy rate was 42.2 % (21.2 % from fresh cycles plus 21.1% from
volume (OV) and ovarian vascularization indexes (OVI). Stored volumes thawed cycles).
were analyzed by the VOCALTM program. Mean greyness-MG, vascularization Conclusions: In our experience oocyte cryopreservation is an option to avoid
index, flow index and vascularization flow index were serially calculated supernumerary oocytes disposal in centers where embryo cryopreservation
throughout the follicular phase of ovarian stimulation. Serum FSH, LH and is not allowed. Although the survival rate is not high when all the
insulin were also checked. Ovulation was induced by hCG administration. supernumerary oocytes are frozen, the one that do survive give rise to
Results: We observed conflicting results in PCOS women as OVI appeared embryos with acceptable implantation rate.
positively related to FNPO rather than OV or FSH /LH levels. In the presence
of FNPO > 20 OVI were all significantly higher in PCOS women, than in
normal ones, in every step of stimulated cycles independently from pituitary P.061 – OOCYTE CRYOPRESERVATION. CHILEAN CENTER EXPERIENCE
down regulation, whereas in the presence of FNPO > 12 but < 20 Carolina Ortega, Isabel Carrasco, Marina,Ríos, Veronica Sáez, Patricio
(consensus standard) OVI were lower in PCOS women following pituitary Donoso, René Salinas, Rodrigo,Enriquez, Patricio González. Reproductive
suppression. MG values were similar in both groups but total stromal area Medicine Unit.ClÌnica Alemana Stgo.U del Desarrollo.Stgo,Chile., Santiago,
was significantly increased in PCOS. Normoandrogenic ovulatory women Region Metropolitana, Chile.
needed a higher amount of -follitropin to get the same PFBF vascularization Introduction: Oocyte cryopreservation (OC) has become an interesting
indexes than the PCOS ones. Insulin resistance appeared positively related alternative in countries or institutions in which embryo freezing is forbidden.
with OVI increasing. In addition, this technique can be offered to women at risk of premature
Discussion: PCOS is a heterogeneous disease. FNPO better relate with ovarian failure (oncologic treatment), or those who have decided to delay
ovarian vascularization and angiogenesis during COH than FSH/LH levels or motherhood. Our OC program started in October 2006 under the framework
OV. Women with a total FNPO > 20 are at higher risk of ovarian of a limited number of oocytes to be inseminated since embryo
hyperstimulation. Insulin resistance may play a role in ovarian hyper cryopeservation is not allowed. All surplus mature oocytes are
response in PCOS. cryopreserved using slow-freezing (SF) protocol. The aim of this study is to
show our results from frozen-thawed cycles including oocyte survival,
fertilization, implantation, embryo quality and clinical pregnancy rates.
P.060 – CRYOPRESERVATION OF SUPERNUMERARY OOCYTES. RESULTS
Materials and Methods: We included 23 patients (25 frozen-thawed cycles)
Isabel Margarita Carrasco1, Carolina Ortega1, Marina Ríos1, Verónica Sáez1, who had cryopreserved supernumerary oocytes from a previous
Patricio Donoso1, René Salinas1, Rodrigo Enríquez1, Patricio González1. unsuccessful fresh cycle (November 2006 until October 2008). SF was
1
Reproductive Medicine Unit., Clínica Alemana de Santiago. Universidad del performed in all cycles using 1,5 M propanediol and 0,2 M sucrose in a
Desarrollo. Santiago, Chile. Choline based Medium. Oocytes were loaded into plastic straws and
Introduction: Oocyte cryopreservation is a new option for countries and/or transferred to a Cryo Bath Freeze Control. Decreasing concentrations of
centers where embryo cryopreservation is not allowed. This technique propanediol and sucrose made thawing procedure. During the thawing
represents a tool to preserve supernumerary oocytes after ovarian cycle, patients were followed up on a natural cycle until the leading follicle
hyperstimulation, avoiding the medical risks and costs of a new cycle. The reached at least 17 mm. hCG (5000 IU) were administered to enable proper
aim of this report is to communicate the first results in our country of an timing with the thawing process. Oocytes were thawed after 36 hours in
oocyte slow freezing cryopreservation protocol in IVF patients, freezing all straws of 5 oocytes. ICSI was done 2 hours after thawing. Embryo transfer
the mature supernumerary oocytes. Oocyte survival, fertilization, embryonic was performed on days 2 or 3 of development. The number of oocytes to be
development, implantation and pregnancy rates are analyzed. inseminated was decided considering women’s age and diagnosis of
infertility. A ßhCG test was performed on day 15 after ICSI and US 3 weeks
Material and Methods: Supernumerary oocytes from patients who did not
after. Micronised vaginal progesterone as luteal support was started on the
get pregnant after a conventional IVF cycle and had at least 3 oocytes
day of ICSI.
available for cryopreservation, from November 2006 to October 2008. Slow
freezing technology was performed. Oocytes were denudated and placed in Results: A total of 524 supernumerary mature oocytes from 56 cycles (54
a choline based medium with 1.5M PrOH for 10 minutes, followed by 15 patients) were frozen with SF technique. Of these, 187 oocytes (25 cycles)
minutes on the same medium plus 0.2 M sucrose. Oocytes were loaded in were thawed and 19 cycles (17 patients) were transferred. The mean age
plastic straws and transferred into an automated Cryo Bath Freeze Control. was 33.4 ± 4.4 years (24-42). The survival rate per thawed oocytes was
Temperature was decreased gradually from 22ºC to -6ºC and seeding was 41,2% (77/187). The injected oocytes were 3,6 ±1,4 and the transferred
performed. After 10 minutes temperature was decreased in 3 steps (-32ºC, embryos were 2,3 ± 1,2. The fertilization, implantation and clinical
-40ºC and free fall). Straws were placed in liquid nitrogen at -196 ºC. pregnancy rates per thawed oocytes were 61%, 14% and 24% respectively.
The embryonic development was 93,6% with 34,1% good quality embryos.
For oocyte thawing the cryoprotectant was removed at room temperature by
The number of live birth was 4 (16%), three healthy children, one preterm
serial dilutions using 0.2M, 0.1M and 0M sucrose solutions. Oocytes were
delivery at 24 weeks who did not survived and three spontaneous abortions.
cultured in Cook Fertilization Medium at 37ºC in an atmosphere of 6% CO2 in
No multiple pregnancies were recorded. The cumulative pregnancy rate per
air. ICSI was performed 2-3 hours after thawing. The number of
transfer was 46,4%.
microinjected oocytes per patient (1 to 5) did not deferred from fresh cycles
April 19-22, 2009 90

Conclusions: The present study showed the results of thawing cycles that Supported by Oncofertility Consortium NIH 1 UL1 RR024926 (R01-
did not achieve pregnancy in fresh cycle, hence when patients who become HD058294, PL1-EB008542), NCRR-RR00163 and U54 HD55744.
pregnant in the fresh cycle choose to thaw their oocytes, the cumulative
pregnancy rate may rise.
P.063 – CORRELATION BETWEEN BODY MASS INDEX AND EMBRYOLOGY
The survival rate was 42,6%, and reflects the survival of all cryopreserved OUTCOMES IN ART PATIENTS AFTER TRIGGERING FINAL FOLLICULAR
mature oocytes, no matter the oocyte quality before freezing, maternal age MATURATION WITH 250 MICROGRAMS OF RECOMBINANT HCG
or response to stimulation protocols.
Kathleen Peters1, James Catt2, Diego Ezcura3; 1Merck Serono, Sydney,
Oocyte cryopreservation is an option to improve pregnancy rates in those Australia; 2Monash IVF, Melbourne, Australia; 3Merck Serono SA Geneva,
centres in which embryo freezing is not allowed or for those couples who Switzerland.
reject embryo cryopreservation.
Introduction: The objective of this retrospective observational study was to
evaluate if 250 mcg recombinant human chorionic gonadotropin (r-hCG)
P.062 – CUMULUS OOCYTE COMPLEXES FROM SMALL ANTRAL FOLLICLES achieved similar embryology outcomes in women with different body mass
DURING THE EARLY FOLLICULAR PHASE OF SPONTANEOUS CYCLES indexes (BMI).
IN RHESUS MONKEYS CAN EXPAND AND YIELD OOCYTES CAPABLE Materials and Methods: Two hundred ninety two (292) assisted reproductive
OF MATURATION IN VITRO technology (ART) patients eligible for intracytoplasmic sperm injection
Marina C. Peluffo1, Richard L. Stouffer1,2, Jon .D. Hennebold 1,2, Mary B. (ICSI), treated at Monash IVF (Melbourne Australia) between September
Zelinski1. 1Division of Reproductive Sciences, Oregon National Primate 2007 and December 2007, were included in the analysis. Women were
Research Center; Beaverton; 2Department of Obstetrics and Gynecology, treated with GnRH agonist for mid-luteal down-regulation. Controlled
Oregon Health & Sciences University, Portland; OR, USA. ovarian stimulation was initiated with 150 - 450 IU recombinant human FSH
Introduction: Cryopreservation of the ovarian cortex is one experimental (r-hFSH) depending upon patient etiology. Final follicular maturation was
option for restoring fertility in cancer survivors. This strategy relies on the induced with 250 mcg r-hCG (Ovidrel®) when at least two follicles had
development of primordial/preantral follicles post-thaw in vivo or in vitro to reached 18 mm diameter. Oocyte retrieval was scheduled 38 hours following
yield mature oocytes for subsequent in vitro fertilization of intracytoplasmic r-hCG administration. ICSI was performed on all metaphase II oocytes.
sperm injection. However, there are many small antral follicles (SAF) in the Syngamy was assessed at 24 hours post-insemination (hpi), day 2 between
ovarian medulla that could be a potential source of oocytes for in vitro 42 – 44 hpi, day 3 at 64 – 68 hpi. Blastocyst transfer occurred on day 5
maturation (IVM) in tissue not used for cryopreservation. The aim of this post-OPU. Patients were stratified according to their BMI: 17-20 kg/m2
study was to determine the minimal diameter of an antral follicle from (Group 1), 21-25 kg/m2 (Group 2), 26-29 kg/m2 (Group 3), 30-48 kg/m2
spontaneous menstrual cycles in macaques that could yield a cumulus- (Group 4). Data was analyzed utilizing JMP 7.0 software (SAS Institute).
oocyte complex (COC) capable of expansion and oocyte maturation in vitro. Continuous variables were expressed as mean ± standard deviation and
categorical variables were expressed as percent. ANOVA was utilized to
Materials and Methods: Ovaries were removed from adult rhesus monkeys analyze continuous variables and Chi Square for categorical data; statistical
(n=6) during the early follicular phase (days 3-4) of spontaneous cycles. significance was established at p<0.05.
SAF were dissected from the ovarian medulla after collagenase treatment.
The COCs from healthy SAF (devoid of dark oocytes or granulosa cells) were Results: Of the four BMI groups (G1 – G4), 36 patients were assigned to G1,
extracted (n=87). SAF were divided into five groups according to their 96 to G2, 32 to G3 and 34 to G4. There were no significant differences
diameter; Group 1: < 0.5 mm; Group 2: 0.5-0.99 mm; Group 3: 1.0-1.49 among the different age-groups, which ranged from a mean of 33.8 (±3.6)
mm; Group 4: 1.5-1.99 mm; and Group 5: 2.0-2.5 mm. Individual COCs to 34.8 (±4.4). Likewise there were no significant differences among the
from each group were cultured for 48 h in TALP (Tyrode’s, albumin, lactate, four groups in the mean daily dose (in IU) of FSH (G1=262±119,
pyruvate) alone, TALP + follicle stimulating hormone (FSH) + luteinizing G2=258±110, G3=244±115, G4=290±108). No significant differences among
hormone (LH) (n=46 COCs), SAGE (SAGE®, CooperSurgical, Inc.) alone, or the four groups were noticed in the total number of oocytes retrieved
SAGE + FSH + LH (n=41 COCs). Images of COCs were acquired at collection (G1=9.6±6.0, G2=10.8±5.5, G3=10.9±6.4, G4=9.1±6.5) as well as the
(0 h), 24 and 48 h post-treatment to assess cumulus cell expansion. At 48 number of metaphase II oocytes available for injection (G1=7.8±5.2,
h, oocyte maturation and diameter were measured after treatment of COCs G2=8.8±5.0, G3=8.8 +-4.9, G4=6.7±4.5). In terms of embryo characteristics,
with hyaluronidase. there were no differences among the groups in the number of 2 pronuclear
(PN) embryos (G1=5.6±4.2, G2=5.8±3.8, G3=5.3±3.3, G4=4.6±4.0); the
Results: Of the total COCs collected, the majority distributed into Group 3 number of embryos transferred (G1=1.1±0.3, G2=1.1±0.4, G3=1.2±0.4,
(69%), with fewer in Groups 1 (2%), 2 (14%), 4 (9%) and 5 (6%). In all G4=1.3±0.4) or in the number of embryos cryopreserved (G1=1.5±2.5,
groups, cumulus-oocyte expansion was seen 24 and/or 48 h post-treatment. G2=1.3±1.6, G3±0.9±1.2, G4=1.0±1.2).
There were no differences in the percentage of oocytes that resumed
maturation to metaphase I (MI) or continued maturation to metaphase II Conclusions: The observations of this study need to be further tested in a
(MII) at 48 h in vitro between media alone and media + gonadotropins, so prospective randomized clinical study.
data were combined within media. Oocyte maturation was not observed in
Group 1 in either media. In TALP, the percentages of MI oocytes observed in P.064 – OVARIAN HYPERSTIMULATION SYNDROME – DO ASIANS DIFFER
Groups 2, 3 and 4 were 20% in each and were similar to those seen in SAGE FROM THE WESTERNERS?
(14% and 20% in Groups 2 and 3, respectively). A greater proportion of
oocytes achieved MII in both media. The percentage of MII oocytes Hemashree Rajesh1, Stephanie Fook-Chong2, Su Ling Yu1. 1Department of
obtained were not different (p >0.05) when cultured in either TALP or SAGE Obstetrics and Gynaecology Singapore General Hospital, 2Department of
in Groups 2 (40 vs. 29%), 3 (31 vs. 16%) and 4 (40 vs. 33%). The diameter Clinical Research, Singapore General Hospital; Singapore.
of MII oocytes was similar regardless of SAF diameter, and did not differ Introduction –We aim to to identify the variables associated with ovarian
between TALP (108 ± 2 μm, mean ±SEM) and SAGE (107 ± 6 µm). hyperstimulation in the Asian patients profile and compare them with
Conclusions: These data indicate, for the first time in primates, that western standards. In addition characteristics between patients with a
cumulus-enclosed oocytes derived from healthy SAF at least 0.5 mm in polycystic ovary( PCO) on ultrasound and those without one( NPCO) were
diameter obtained during the early follicular phase can meiotically mature in compared.
vitro. The competence of these oocytes to undergo fertilization after IVM Materials and Methods - This was a retrospective case analysis of in vitro
with subsequent embryonic development is under investigation. Thus, the fertilisation(IVF) records of patients at the Centre for Assisted Reproduction
cohort of SAF present in the ovaries of spontaneous menstrual cycles prior in a tertiary restructured hospital at Singapore, who developed moderate or
to selection of the dominant follicle can provide COCs as an additional severe ovarian hyperstimulation syndrome( OHSS) during IVF stimulation. A
source of oocytes for IVM and fertility preservation. Whether COCs obtained total of 79 patients identified over 5 years were subdivided into early onset
from SAF outside of the early follicular phase are also suitable for IVM and late onset hyperstimulation. Patient characteristics, ultrasound
remains to be determined. features, gonadotropin doses, stimulation sequelae and response were
91 Geneva
analysed. We used the Rotterdam criteria in the ultrasound recognition of alternative to preserve fertility in those patients in whom conventional
polycystic ovaries. Statistical analysis was done using SPSS.B L E 1 controlled ovarian stimulation is contraindicated.
Results: 63% of the population were Chinese with the rest - a mixture of
other Asian nationalities. The incidence of moderate to severe early OHSS P.066 – PER-FOLLICLE ANTI-MULLERIAN HORMONE SECRETION AND
with an intention to treat was 10.9%( actual incidence 9.1%) with 86.1% PERIFOLLICULAR POWER DOPPLER ANGIOGRAPHY MAY REFLECT
having early OHSS. 63.3% had a PCO picture on ultrasound. QUANTITATIVELY AND QUALITATIVELY THE OVARIAN FOLLICULAR
The mean estradiol level at hyperstimulation in a long cycle was comparable STATUS IN A HIGH-RISK POPULATION OF INFERTILE WOMEN
between a patient with PCO and non PCO( Mean 6009.1 pg/ml, vs Mean Annalisa Salerno, Alfredo Nazzaro. Physiopathology of Human Reproduction
5391pg/ml, p=0.25). The average gonadotropin dose associated with Unit, AORN “G. Rummo” Hospital, Benevento, Italy.
hyperstimulation was also comparable. Gonadotropin doses and maximum
estradiol levels did not vary between the long cycle and antagonist cycle in a Objective: The determination of ovarian reserve continues to be a challenge
woman <35 years with PCO(Mean 204.2 iu vs 168 iu). and we are at the continuous research of affordable markers helpful to
predict cycle cancellation and quantitatively/qualitatively poor response to
There was no difference in BMI or severity of hyperstimulation between ovarian stimulation. There are evidence that antimüllerian hormone (AMH) is
patients with or without PCO ( p=0.600). However hyperstimulation involved in both the control of primordial to primary follicle growth and their
showed a tendency to be more severe in a PCO patient with a low BMI. responsiveness to FSH administration and that the acquisition of oocyte
Hyperstimulation was attributed to a high starting dose in 72.1% of patients developmental competence is secondary to perifollicular vascularization
and to a 50iu step up in 23.5%. (PV) and intrafollicular O2 content. In this study we have investigated the
After stimulation, ultrasound on day 5- 7, showed a higher follicle count in relationship between per-follicle AMH levels, PV, ovarian follicular status
a patient with PCO than in a non PCO (p=0.001, mean 28 versus 20). This and oocyte quality during controlled ovarian hyperstimulation (COH) in a
correlated with a significantly high initial antral follicle count in the PCO high risk population of infertile women.
group( median 22.5 vs 10.5, p=0.003). At peak stimulation, there were Materials and Methods: a total of 162 ICSI patients at high risk of cycle
more large follicles(> 14) (p=0.010) and a trend towards more intermediate cancellation due to raised FHS, previous poor response and/or age ≥ 40
follicles(p=0.079) in the PCO group. There was no significant difference in years were studied. All of them underwent a cycle day 3 basal measures of
oocyte retrieval between the two groups. The number of follicles did not FSH and AMH, ovarian blood flow was evaluated by three dimensional
relate to the grade of OHSS. powerDoppler angiography (3DpDA) and a total antral follicles count (tAFC)
Severe OHSS presented significantly later on day 4( p= 0.004) after oocyte was obtained from 3D ovarian volumes in both ovaries by using a GE730
retrieval than moderate ovarian hyperstimulation(day 2). Prophylactic Voluson machine. Basal ovarian blood flow was evaluated by the VOCAL
albumin helped to reduce severe hyperstimulation. 43 patients had an program. tAFC was obtained by the NICHE program. PV was quantified by
embryo transfer and there was a 25% incidence of pregnancy. 90% of seriate 3DpDA measurements during ovarian stimulation and blood flow
patients who presented with late OHSS were pregnant with a 50% incidence indexes were off line calculated by the VOCAL program set to color
of twins. rendering mode. On the day of oocyte retrieval serum samples and follicular
Conclusions – Gonadotropin doses at stimulation should probably conform fluids from three follicles < 12 mm and three follicles between 17 and 21
to the western standards of 150iu in women less than 35 years and minimal mm in diameter were collected and tested for AMH, E2 and Progesterone
increases of 37.5iu should be considered at step-up. A total follicle count of (P4) contents. Oocyte morphology, fertilization rate and embryo quality were
more than 20 on day 5- 7 scan may be predictive of a subsequent evaluated.
hyperstimulation. Prophylactic albumin should be considered to reduce the Results: Intrafollicular AMH levels were more than double in the follicle ≤12
incidence of hyperstimulation. Transfer should be abandoned in the mm respect to the largest ones. Per-follicle AMH levels appeared to be
presence of high estradiol levels ( > 5200pg/ml) or when the total number related to day 3 tAFC, number of follicles ≥ 12 mm and number of retrieved
of intermediate and large follicle count exceeds thirty on the day of oocyte oocytes. High intrafollicular levels of P4 were inversely related to the AMH
retrieval or when the oocyte retrieval exceeds nineteen eggs. ones in both follicular classes. Per-follicle AMH levels appeared inversely
Hyperstimulation may be more severe as the BMI decreases and late ovarian related with requirement of FSH units during ovarian stimulation. PV indexes
hyperstimulation must prompt a look out for twins. Variables seem to be appeared to be unrelated to follicle diameters and ovarian volume but there
similar in the Asian population as compared with the west. was a positive correlation with AMH levels (p<0.05). tAFC did not interfere
with blood flow indexes. Cycle cancellation rate was correlated either with
AMH levels (p<0.05) either with stromal blood flow (p<0.002), whereas tAFC
P.065 – IVM EXPERIENCE IN MEXICO, INITIAL EXPERIENCE alone did not (p>0.05). Women with basal AMH levels <1.60 ng/ml showed
Luis Arturo Ruvalcaba, Alejandro Chavez-Badiola, Rocio Martinez, Jose lower quality oocytes (dark central granulation, increased viscosity,
Medina, Martha Garcia. Instituto Mexicano de Infertilidad, Zapopan, Jalisco, aggregation of smooth endoplasmic reticulum, increased vitelline space)
Mexico. compared with the ≥ 1.60 ng/ml ones. The number of competent oocytes
dropped to zero in case of undetectable basal stromal blood flow
Background: Results for in-vitro maturation (IVM) are improving and
indipendentely from AMH levels (p<0.001). tAFC did not seemed to affect
becoming a reality in several countries. Its low cost and low risk profile is
oocyte quality p(>0.05) but the number (p<0.05). Undetectable stromal
very attractive and probably should be considered as a treatment option in
blood flow but not AMH levels affect fertilization rate and embryo grading
developing countries. In this communication we present our experience with
(p<0.001).
IVM cycles in a Mexican IVF unit.
Conclusions: Per-follicle AMH levels and 3DpDA may be used as prognostic
Methods: Patients with polycystic ovaries were recruited to our IVM biomarker(s) of oocyte competence. The routinary use of 3DpDA to detect
programme. Collected oocytes were matured as described by Chian. basal stromal blood flow combined with AMH levels seems to be superior to
Reported results are given in percentages of maturation and live-birth per AMH alone to predict both oocyte quality and developmental competence.
series. Cycle day 3 basal ovarian blood flow and anti-Mllerian hormone (AMH) level
Results: We achieved an overall 82% maturation rate from MII and GV are useful tools to predict diminished oocyte quality and developmental
oocytes. Although some patients are waiting to complete their treatment incompetence in infertile women.
cycles, this far we have only achieved a live-birth. In this case the patient
had to undergo vitrification of IVM oocytes due to personal reasons. A
single intrauterine pregnancy was identified at seven gestational weeks
(25% implantation rate). A healthy baby girl was delivered at 38 gestational
weeks.
Conclusions: IVM is a promising tool in the field of ARTs and although we
are still looking to improve our pregnancy rates, we support that IVM
followed by oocyte cryopreservation is, according to our results, a viable
April 19-22, 2009 92

Results: 70 patients underwent aspiration of follicles. 14 of these had

P.067 – ASYNCHRONOUS FROZEN EMBRYO TRANSFERS – DOES IT AFFECT unexpected ovulation. Another 11 had an “empty” large follicle and from 45
RESULTS? women a mature oocyte was retrieved. In cycles with EFS the FF-
concentration of progesterone were lower than in cycles with a mature
Tarique Salman, Amanda Tozer, Talha,Al-Shawaf, Luca Sabatini, Ariel oocyte retrieved (9.5 ± 2.2 vs. 13.5 ± 1.1) µg/ml, with p = 0,037 despite the
Zosmer. Centre for Reproductive Medicine, St. Bartholomew’s Hospital, fact that US- criteria were met.
Barts and the London NHS Trust, West Smithfield, London, UK.
Conclusion: Cycles with EFS exhibit different hormonal profiles than cycles
Introduction: In frozen embryo transfer (FET) cycles the transfer is usually with mature oocytes retrieved.
planned so that embryos’ and the endometrial development will be
synchronous [i.e. day 2 (D2) embryos are transferred on D2 of the luteal
phase etc.). Little information is available as to whether asynchrony may P.070 – INCIDENCE OF OVARIAN HYPERSTIMULATION SYNDROME IN
affect the outcome. PATIENTS AFFECTED BY POLYCYSTIC OVARY SYNDROME
Materials and Methods: Data for FET cycles (1.1.2005 and 31.12.2007) were UNDERGOING IN VITRO FERTILIZATION: A PRELIMINARY
retrospectively collected from our computerised database and patients’ RANDOMIZED STUDY
notes. In natural cycle FET (NC-FET) ovulation (day 0) was consider to occur Alessandra Tirelli, Simone Giulini, Antonio La Marca, Francesca Tortolani,
the day after a positive urine LH test. In hormone replacement cycles (HRT- Susanna Xella, Daniela Tagliasacchi, Tiziana,Marsella, Annibale Volpe.
FET) day 0 was considered the day when progesterone was started. Department of Obstetrics and Gynecology, University of Modena, Modena,
Results: 760 cycles were included: 666 synchronous (control), 94 Italy.
asynchronous (study). The clinical pregnancy (CPR) and live birth (LBR) Introduction: Several ovarian stimulation protocols have been proposed over
rates per transfer were 18.5% and 14.9% vs. 17.0% and 15.9% respectively the years for patients affected by polycystic ovary syndrome (PCOS)
[(not significant (NS)]. The LBR for NC-FET or HRT-FET were 17.7% and undergoing in vitro fertilization (IVF); however the optimal stimulation
10.9% (control) vs. 16.8% and 12.5% (study) (NS). In the study group the protocol is still under debate.
LBR for D2 embryos transferred on D3 was 16.6% (NC-FET – 17.1%; HRT- It is expected that a satisfactory number of oocytes can be recovered
FET – 14.2%) and that for D3 embryos transferred on D2 was 12.5% minimizing the risks of ovarian hyperstimulation syndrome (OHSS). OHSS
(NC-FET – 14.2%; HRT-FET – 11.1%). In the control group the LBR for D2 is an exaggerated response to ovulation induction therapy and such
transfers was 15.2% (NC-FET – 17.8%; HRT-FET – 11.5%) and that for D3 complication should always be accounted for in PCOS patients.
transfers was 10.6% (NC-FET – 16.6%; HRT-FET – 4.3%). There was no
significant difference between any of the LBRs in the control and study In the present study we compare the incidence of OHSS in PCOS patients
groups. undergoing IVF, treated with Gonadotrophin Releasing Hormone (GnRH)
agonist protocol or with GnRH antagonist protocol.
Conclusions: A one day asynchrony does not seem to affect the treatment
outcome. The effect of a longer asynchrony needs further assessment. Material and Methods: PCOS was diagnosed according to the Rotterdam
ESHRE/ASRM consensus on diagnostic criteria (2004). Fifty-two patients
affected by PCOS were enrolled and randomly assigned to two different
P.069 – ULTRASOUND MONITORING AND HORMONAL PROFILES IN SERUM ovarian stimulation protocols. GnRH-agonist group (Group 1; 29 cases)
AND FOLLICULAR FLUID FOR UNSTIMULATED IVF-CYCLES IN received Leuprorelin 3.75 mg (Enantone 3,75; Takeda) in the midluteal
RELATION TO “EMPTY FOLLICLE SYNDROME” phase and stimulation with 150 IU rFSH/day (Gonal-F; Serono) at least 15
Mikael Tang-Pedersen, Lars Westergaard. Fertility Clinic, Odense University days after agonist injection. GnRH-antagonist treated patients (Group 2; 23
Hospital; Faculty of Health - Institute of Clinical Research, University of cases) received 150 IU of rFSH/day starting on day 2-5 of menses,
Southern Denmark, Odense, Denmark. followed by Cetrotide 250 μg/day (Orgalutran, Organon) when the lead
follicles were 13–14 mm in mean diameter.
Introduction: Over the years one observation remains constant in
unstimulated / natural cycle IVF. That is the approx. 20 % of so called OHSS occurrence was diagnosed and classified according to a standardized
“empty follicles”. Empty follicle syndrome (EFS) has been debated, but since clinical criteria. The incidence of OHSS was compared among the two
it seems a constant finding a deeper understanding of the follicular groups. Additionally, pregnancy rate, dose of gonadotropins admnistered,
maturation process is desirable. stimulation time, number of oocytes retrieved, number of embryos
transferred and number of “good embryos” on day 3 were assessed and
In this study we investigate hormonal levels in follicular fluid and serum for compared among groups.
unstimulated IVF-treatment of ± oestradiol-primed patients from start of the
cycle till day of oocyte pick-up (OPU), along with ultrasonic measurements Results: Six patients from Group 1 developed a mild OHSS during
of follicle size and number and endometrial thickness. stimulation and the treatment was thus suspended. No treatment was
suspended in Group 2. Mild OHSS after embryo-transfer (ET) developed in 7
The aim is to explore possible correlations between endocrinology, patients from Group 1 (24,1%) and in 3 patients from Group 2 (13%).
ultrasound monitoring and failed cycles due to “EFS”. Moderate OHSS was observed in 3 patients (10,3%) in Group 1 and in 0
Materials And Methods: 70 women under the age of 37, referred to IVF- patients in Group 2. No patient developed severe OHSS. Differences
treatment due to tubal, male or unexplained factor and with regular cycles between groups were statistiscally significant according to one-way analysis
from 26-34 days and FSH <15 U/L were included in this project as a of variance (ANOVA) (P<.005).
retrospective analysis. Pregnancy was achieved in 8 patients in Group 1 (pregnancy rate per
Ultrasound (US) examination and blood samples were scheduled beside day cycle=27.6%; pregnancy rate per ET=34.8 %) and in 7 patients in Group 2
of OPU to early, mid and late follicular phase until the appearance of a (pregnancy rate per cycle=30.4%; pregnancy rate per ET=30.4%). Mean
dominant follicle > 16 mm, with a corresponding endometrium > 8 mm. An total amount of gonadotropin units administered per patient in Group 1 was
hCG-injection of 6500 IU is administered 34 hours prior to OPU. The 1949 ± 95 IU and 1480 ± 169 IU in Group 2, (statistically significant; P<.01).
aspiration of the mature follicle provides follicle fluid for measuring levels of The number of oocytes retrieved were significantly higher in Group1 (12.8 ±
AMH, progesterone, oestradiol, androstendion and testosterone. The mature 1.1 vs. 7.8 ± 0.7, respectively; P<.003). Stimulation time; number of
oocyte is fertilized and transferred 2 days post-OPU. embryos transferred and number of “good quality” embryos did not
statistically differ in the two groups.
35 of the 70 women had oestradiol-priming, 2 mg twice daily (Femanest®,
Sandoz), in 3-10 days in order to predict most likely time for LH-peak and Conclusions: GnRH antagonist protocol is associated to lower incidence of
hence avoid premature ovulation as described in earlier work by deZiegler et OHSS in PCOS patients, without reducing pregnancy rate.
al.
Delaying intermenstrual FSH-rise until a pre-defined day of lowering serum
oestradiol is the feed-back mechanism behind this option.
93 Geneva
pregnant. Regardless of the groups, over 50% patients that gave up on their
P.071 – EFFECTS OF ESTRADIOL/OOCYTE RATIO ON THE OUTCOME OF treatment gave up within 6 months and the other hand, 90% of patients that
CONTROLLED OVARIAN STIMULATION FOR ASSISTED REPRODUCTIVE were introduced to birth facilities have become pregnant within 2 years.
TECHNOLOGY CYCLES WITH GONADOTROPIN RELEASING HORMONE Consequently, we found that it is very important to support infertility
AGONIST patients with continued treatment, especially during the early stage of their
treatment.
Esra Tonguc, Turgut Var, Muammer Dogan, Leyla Mollamahmutoglu.
Department of Reproductive Endocrinology, Zekai Tahir Burak Women’s
Health Research and Education Hospital, Ankara,Turkey. P.073 – CLINICAL EFFICACY OF A NOVEL EVALUATION METHOD WITH
Introduction: The aim of this study was to evaluate the effect of estradiol MEASUREMENT OF EMBRYO RESPIRATION ACTIVITY USING A
level on the day of HCG(peak E2)/oocyte ratio on outcome of IVF using a SCANNING ELECTROCHEMICAL MICROSCOPY
gonadotropin releasing hormone agonist (GnRH-agonist) based controlled Takafumi Utsunomiya1, Yoko Kumasako1, Kaori Goto1, Megumi Koike1,
ovarian stimulation (COH) for assisted reproductive technology (ART ) cycle. Masaki Yokoo2, Hiroyuki Abe3. 1St-Luke Clinic, Oita, Japan, 2Tohoku
Materials and Methods: Of the patients who underwent IVF-ET at the University Biomedical Engineering Research Organization (TUBERO),
Department of Reproductive Endocrinology at our hospital between the Sendai, Japan, 3Gradeuate School of Science and Engineering, Yamagata
years 2005-2007, 600 normal and high responders to the first cycle of COH University, Yonezawa, Japan.
with GnRH-agonist were included in the study. Data was obtained from Introduction: Respiration is a useful parameter for evaluating embryo quality
patient records. Patients were designated into 3 groups based on peak as it provides important information about metabolic activity. The scanning
E2/oocyte ratio (Group A:<100 pg/ml, Group B:100-200 pg/ml, Group electrochemical microscopy (SECM) measuring system provides non-
C:>200 pg/ml). A comparison between groups was made regarding ovarian invasive and accurate measurement of the oxygen consumption (respiration
stimulation characteristics, number of oocytes obtained, number of activity) of single human embryos. We have shown that there is correlation
transferred embryos, fertlization and pregnancy rates. between embryo quality and respiration activity in human embryos at the
Results: After the division based on E2/oocyte ratio, in Group C early developmental stage. In this study, for practical purposes, we
(E2/oocyte:>200), the number of obtained oocytes, 2 PN (fertilized oocyte), examined the clinical efficacy for IVF-elected single embryo transfer (eSET)
M2, number of total and transferred embryos was statistically significantly patients using a modified-SECM measuring system.
lower than in the other two groups (p=0.001, 0.001, 0.001, 0.039, 0.045). Material and Methods: A total of 121 IVF-eSET patients gave consent to
Patients in group C had a significantly higher level of E2 on HCG day their cycles being evaluated using a randomized study. In the morning after
(p=0.001). However, there was no statistically significant difference between 2-3 days culture following the conventional IVF or ICSI procedure, we
groups regarding number of high quality embryos, fertilization and evaluated the embryo quality using a morphological method, and the oxygen
pregnancy rates consumption of individual embryos was measured using a modified SECM
Conclusions: The E2/oocyte ratio has no predictive value in determining system. All cycles had two or more embryos that appeared reasonably good
pregnancy rates for normal and high responders to IVF cycles using COH morphologically on ET day (ex. better than 6cell grade 3 by Veeck’s method
and GnRH-agonist. on Day3). Each embryo was transferred into a plate filled with medium. A
Pt-microdisk electrode was lowered into the solution, and its tip potential
was held at -0.6V to monitor the local oxygen concentration.
P.072 – INFERTILITY PATIENT’S MENTAL HEALTH CONDITION USING THE A single embryo, based on morphological evaluation (ME) or combing
CORNELL MEDICAL INDEX morphological evaluation and measuring respiration activity (ME+MR), was
Keiko Ueno, Michiyo Sashiyama, Takafumi Utsunomiya. St. Luke Clinic, Oita, elected by either decision and transferred into the patient’s uterus.
Oita, Japan. Results: In this study, seventy nine patients’ cycles (reasonably good
Purpose: In 1997, the mental health condition of infertility patients in our morphologically on ET day) were devoted (61 cycles of ME eSET and 18
clinic was evaluated using the Cornell Medical Index. “As a result, the cycles of ME+MR eSET). Pregnancy rate following of eSET by ME
patients with a high neurosis tendency did not continue medical treatment (conventional election) and ME+MR (novel method) was 39.3% (24/61) and
as well as attending clinic regularly. The period was shorter than the others.” 50.0% (9/18), respectively. This result suggests a clinical efficacy for
embryo quality evaluation by measurement of respiration activity.
Based on the above result, infertility patient’s mental health condition was
researched once more using the Cornell Medical Index when they visited our Subsequently, among all the cycles that had two or more embryos
clinic for the first time and when we introduced them to birth facilities representing exactly the same excellent morphology by Veeck’s method (ex.
during their pregnancy. 8cell grade 1), the pregnancy rate in cycles (n=21) that transferred a single
embryo which was elected based on only morphology by examination under
Object and Method: From February 2001 to May 2003, Cornell Medical
the microscope was 38.1%, (8/21). Intriguingly, cycles (n=20) that
Indices were distributed to 464 female patients when they visited our clinic
transferred a single embryo which was elected based on morphology and
for the first time (the response rate was 93%; 433 patients) . From October
respiration activity had a higher pregnancy rate: It was 60.0% (12/20).
2003 to October 2004, Cornell Medical Indices were distributed to 238
female patients that became pregnant after treatment, at that time they were Until now, 6 patients of eSET based on ME+MR resulted in a singleton
introduced to birth facilities (the response rate was 77%; 184 patients). Our pregnancy and the birth of normal healthy babies (4 male and 2 female)
examiner handed the Cornell medical Index to patients and they were weighing 2520-3142g after 37-40 weeks’ gestation.
collected later. Conclusions: It is very difficult to judge which embryo has the most viability
Result: 60% of the patients, belonging to group 4 (high neurosis tendency) of all in eSET. Selecting one viable embryo using only the morphological
when they visited our clinic for the first time, discontinued treatment, and method seems less objective. The results of this study support the
90% of these patients gave up treatment within 6 months. A significant hypothesis that measuring embryonic respiration provides additional and
difference was seen between group 4 and the other groups. Through this valuable information about the embryo quality. It is indicated that the
research, we can say that the patients in group 4 did not continue treatment highest pregnancy rate will come to fruition by electing the best embryo by
regularly for a long period of time (p< 0.05). Regardless of the groups, over combining morphology evaluation and respiration activity evaluation in IVF-
50% of the patients that discontinued their treatment gave up within 6 eSET. It is hoped that this new, novel method will prove to be more effective
months. If they belong to group 4, and they continued treatment regularly for IVF treatment.
over 6 months, they would have become pregnant within 2 years. The result
of the Cornell Medical Index at the time of introduction to birth facilities, the
rate of group 1 (the healthiest group) was significantly increased in
comparison with their first visit to our clinic (p < 0.01).
Conclusion: With reference to p<0.05, compared with the other normal
groups, high rate of group 4 patients gave up treatment before they got
April 19-22, 2009 94

from the leading follicles and the small follicles; Group 2 (n=75 cycles):
P.074 – THE INFLUENCE OF AGE ON THE BLASTOCYST GROWTH Transferred embryos derived from the oocyte from the small follicles only.
Yuji Fujino, Eiko Wakimoto, Naomi Iida, Mayuko Hori, Kouji Koike, Megumi Results: The rate of clinical pregnancy in Group 1 (43.0%) was higher than
Hoki, Yuka Koma, Ayako Yokoyama. Fujino Ladies Clinic. Suita, Osaka, that in Group 2 (37.3 %) but there was no significant difference between
Japan. groups. Implantation rate were not significantly different between Group 1
(17.4%) and Group 2 (16.3 %).
Introduction: The aim of this study is to examine the results of blastocyst
culture and transfer depending on female age for establishing an efficacy Conclusion:These results indicate that the clinical outcome of embryos
strategy for overall reproductive outcome. produced from the oocytes derived from the small follicles is not affected by
the presence of the dominant follicle. These results also suggest that the
Materials and Methods: Clinical evaluation of growing rates to the blastocyst optimal HCG injection time is the size of leading follicles reached to 12-14
stage and pregnancy rates of blastocyst transfer from February 1999 to mm in diameter.
November 2004.Total 2593 cycles were performed oocytes pick-up and
blastocyst culture. Ovarian stimulation was performed with Clomiphene
Citrate (CC) and Gonadotropin (hMG) protocol. They received 50mg of CC P.076 – EFFECTS OF BLOOD AND CERVICAL MUCUS ZINC, COPPER, CADMIUM
beginning on day 3 of her cycle, followed by 150 IU of hMG on cycle day 8 AND LEAD LEVELS ON INFERTILITY IN WOMEN
and day 10. When at least one follicles of diameter reached at >18mm, Halil Ilgin, Atilla Yildirim, Hikmet Hassa. Eskisehir Osmangazi University
GnRH analogue was sprayed nasally. Oocyte retrieval was carried out 32 to (ESOGU) School of Medicine, Dept. Ob&Gyn Reproductive Health Center,
33h later using the flare-up effect of GnRH analogue. Oocytes were Eskisehir, Turkey.
inseminated with sperm and cultured in various sequential media. Blastocyst
was transferred under the ultrasound guidance. Clinical pregnancy was Introduction: This study was conducted to find out the effects of blood and
diagnosed by the detection of gestational sac with the ultrasound. As cervical mucus zinc (Zn), copper (Cu), cadmium (Cd) and lead (Pb) levels
statistic analysis, Correlation regression and Chi square analysis were used on infertility in women.
with StatView program to compare clinical pregnancy rate (PR) and growing Materials and methods: Thirty-five infertile and 15 fertile women made up
rate to the blastocyst (GR) between each age. P<0.05 was considered our study group. The infertile women were referred to Reproductive Health
statistically significant. Center of Dept.Ob&Gyn in ESOGU School of Medicine with no male factor
Results: The mean age of 2593 cycles was 38.6 +/- 4.3 years old (ranged as their cause of infertility. Fifteen controls were women attending the
from 30 to 49 years old). The mean number of retrieved oocytes of all cycles Gynecology Clinic with no fertility problems and using no contraception.
was 3.1 +/- 2.0 (ranged from 1 to 10 oocytes). Cycles having blastocysts Whole blood (WB), blood plasma (BP) and cervical mucus (CM) samples
were 1234 cycles; those were 52.5% of all treated cycles. The GR was were collected between menstrual days 14-21 and kept at -20º C. To obtain
72.3% (34 cycles / 47 cycles) in 30 years old, 62.7% (104 / 185) in 35 years CM samples Gynetics Medical 4502-B IUI cannulas were used. Zn, Cu, Cd
old, 51.2% (111 / 217) in 40 years old, and 24.7% (22 / 89) in 40 years old, and Pb levels were measured in all samples using Hitachi (180-70) Polarized
respectively. There was significant correlation between the growing rates to Zeeman Atomic Absorption Spectrophotometer in the Dept. of Chemistry of
the blastocyst stage and age (P<0.01). The average number of transferred ESOGU Faculty of Science and Letters. Statistical analysis was made using
blastocysts was 1.3 +/- 0.5 per replacement. One hundred twenty eight the SPSS 13.0 for Windows. Ethical approval was obtained from ESOGU
clinical pregnancies were established, which included 12 set of twin. The PR School of Medicine Ethics Committee before initiating the study.
per transfer cycle was 35.0% in 30 years old, 28.3% in 35 years old, 11.3% Results: 1. BP and CM Zn and Cu levels were lower in the infertile group as
in 40 years old, and 11.1% in 45 years, respectively. There were no compared to the controls (p<0.001).
significant differences in PR of over 35 years old (P<0.05).
2. WB Pb levels were higher in the infertile group (p:0.023), whereas CM Pb
Conclusions: Results from this study confirm the expected age-dependent levels did not differ (p>0.05).
decline of the growing rate to blastocyst and IVF pregnancy rates even when
3. WB and CM Cd levels were higher in the infertile group (p>0.05 and
highly selected blastocysts are transferred. The effective treatment for this
p:0.008, respectively).
age related decline maybe a very difficult problem in the recent reproduction
medicine. 4. Cd levels were higher in WB samples and lower in CM in smokers (four
women in each group, eight in total) (p>0.05).
5. Measurements did not differ statistically in women with primary or
P.075 – THE INFLUENCE OF LEADING FOLLICLES IN NATURAL CYCLE IVF/M
secondary infertility (p>0.05).
TREATMENT
6. CM levels of Zn, Cu and Cd increased paralel to the increases of blood
Seong-Ho Yang, Ki- Chul Kim, Chang-Suk Suh; Maria Fertility Hospital,
levels of these elements, whereas this relation could not be found for Pb
Seoul, South Korea.
levels.
Introduction: Recently, it has been reported that successful live births
7. Negative correlation was found between BP Zn and CM Cd, WB Pb and
obtained from natural cycle IVF combined with IVM (natural cycle IVF/M) for
CM Zn, WB Pb and CM Cu (p:0.001, r:-5.50; p:0.008, r:-3.72 and p>0.05, r:-
patients with regular menstrual cycles (Chian et al., 2004; Lim et al., 2008).
2.87, respectively).
However, it is not well documented that how the recruitment of leading
follicles in the ovaries affects the clinical outcome of the treatment cycle. 8. There was negative correlation in CM Zn and Cd and CM Cu and Cd
Therefore, this study was to examine the influence of the leading follicles on (p:0.09, r:-2.44 and p:0.056, r:-2.73, respectively).
clinical outcome of natural cycle IVF/M. 9. We found in our study, mean levels of Zn for BP and CM in women with
Material and Method: A total of 209 patients with regular menstrual cycles no infertility problems as 1560 µg/L and 32.4 mg/L, respectively and mean
underwent 233 completed treatment cycles. When the leading follicles levels of Cu in the same group as 1785 µg/L and 1.32 mg/L.
reached 12-14 mm in diameter, 10,000 IU of human chorionic gonadotropin Conclusions: That BP and CM Zn and Cu levels are lower, Cd levels are
(HCG) was administrated 36-38 hours before oocyte retrieval. The collected higher and WB Pb levels are higher in the infertile women in our study can
mature oocytes were inseminated by ICSI using the husband’s sperm 2 be contributing factors to their infertility, as both Pb and Cd are known
hours later and the immature oocytes were cultured in 1 mL of maturation reproductive toxins. Blood levels of Zn, Cu and Cd reflect their CM levels.
medium. After one day of culture, the cumulus cells of oocytes were That CM Cd levels are high when CM levels of Zn and Cu are low suggests
removed with 0.03% hyaluronidase and mechanical pipetting. In vitro Zn and Cu levels should be optimal for normal reproductive function.
matured oocytes were also inseminated by ICSI. Fertilization was assessed
We thank Temir Ali Demir, Asiye Berber and Zerrin Kaynak from the Dept. of
17-19 hours after ICSI to detect the appearance of two distinct pronuclei
Chemistry of ESOGU Faculty of Science and Letters, and Ahmet Musmul from the
and two polar bodies. Embryo transfer (ET) was performed on day 3 or 4
Dept. of Biostatistics of ESOGU School of Medicine.
after oocyte retrieval. Clinical outcome was analyzed by the transferred
embryos derived from the oocytes retrieved from the leading follicles or not.
Group 1 (n=158 cycles): Transferred embryos derived from the oocytes
95 Geneva
P.082 – PCOD AS RISK FACTOR FOR PRE-ECLAMPSIA: STUDY OF PRE-

P.079 – EFFECT OF LH TREATED OVINE OVIDUCTAL EPITHELIAL CELL CO- ECLAMPSIA,S RISK FACTORS IN SINGLETON PATIENTS IN THE
CULTURE SYSTEM ON MURINE PRE-EMBRYO DEVELOPMENT FATEMIYEH HOSPITAL OF HAMEDAN
Hiva Alipour, Poopak Eftekhari-Yazdi, Abdolreza Rastgarnia, Mohamadreza M. Farimani, R. Niazpor, M. Gharakhani, S. Rabie. Hamedan University,
Baghaban Eslaminejad. Embryology Department, Royan Institute, ACECR, Hamedan, Iran.
Tehran, Iran. Background: Preeclampsia is a major cause of maternal and fetal mortality
Introduction: This study was designed to develop a new co-culture system and morbidity. The incidence of preeclampsia is 2-10% depending on the
and assess the effect of Luteinizing hormone using sequential media to population studied. This disease is priority of WHO but pathophysiology of
promote development and increase the quality of 2-cell Ovine embryos preeclampsia is not absolutely known. With attention of complication
through the 8-16 cell stage to morula and blastocyst stages. preeclampsia,s complications in pregnancy recognize of risk factors is
obligatory for reducing of MMR , NMR and increasing of civil healthy index.
Material and Methods: Monolayers for co-culture were prepared from Ovine
oviduct epithelial cells (OOEC) in DMEM/F12 medium and in vivo-fertilized Methods: Completion of questionnaire throughout direct interview with
2-cell embryos were collected by flushing from superovulated NMRI mice. patients . Kind of study was case/control and each preeclamptic patient was
Co-culture media was treated with hCG as a surrogate for LH because of its matched for age with one control patient and then patients compared with
stability and purity. Embryos were cultured in G1/G2™Ver.5 drops alone and control group. According to the data number of cases was 142 and control
containing LH, as the control groups and on OOEC monolayers in group was 142 too. Details of questionnaire collecting and analyzed with
G1/G2™Ver.5 drops alone and containing LH as the experimental groups. tenth edition of SPSS .
Development and quality rates were determined for all embryos daily and Results: PCOD(history of hirsutism + oligomenorrhea and high BMI before
statistically compared. At the end of the cultivation period, differentially pregnancy) ,UTI , Previous preeclampsia in multiparous women , family
stained trophectoderm (TE) and inner cell mass (ICM) of expanded history of preeclampsia and passive smoking in case group Significantly
blastocysts from each group were examined microscopically. were higher than when compared with control group (P<0.05).
Results: The embryos cultured on an OOEC monolayer in G1/G2™Ver.5 Also high rate of C/S in patients and more consumption of Aspirin , Folic
drops treated with LH had a significantly higher developmental rate than acid, vitamin C & E were seen in preeclamptic group.
those of the group without LH and the control groups (P≤.05). The Conclusion: Some risk factors such as PCOD , UTI , previous preeclampsia ,
blastocysts from the LH treated group, in comparison with the group positive family history and passive smoking may lead to develop
without LH and the control groups, also had a significantly higher mean cell preeclampsia .
number (P≤.05).
Conclusions: These findings suggest that elevated periovulatory LH levels
may promote preimplantation embryo development. These results have P.087 – LARGE HEADED MULTIFLAGELLAR SPERMATOZOA SYNDROME
important implications for assisted reproductive technologies in which co- A. Sellami1, N. Feki1, N. Abid 1, A. Bahloul2, P. Ray3 , T. Rebai1, L. Keskes1.
cultures are used to improve pregnancy rates. Ovine Oviduct Epithelial cell 1
Reproductive Biology laboratory Medical School and 2Male infertility
co-culture system treated by LH could improve in vitro preimplantation Research Unit, Sfax, Tunisia; 3Department Genetics and Procreation,
embryo development both in terms of quality (increasing blastocyst Grenoble, France.
cellularity) and developmental rate.
Introduction: The development of the assisted reproduction technologies,
essentially the intracytoplasmic sperm injection (ICSI), has revolutionized
P.081– CLINICAL OUTCOME OF IN VITRO MATURATION CYCLES: EXPERIENCE the treatment of male infertility. However, the use of this technique in some
OF 717 CASES IN MARIA FERTILITY HOSPITAL particular cases of spermatogenesis disorders may lead to genetically
defective and no viable embryos. Among these types of severe impairment
Jeong-Ho Cha1, Hye-Jin Yoon1, Seok-Yoon Lee1, Won-Don Lee1, Chang-Won of male fertility, the large-headed multiflagellar spermatozoa syndrome is a
Kang2, San-Hyun Yoon3, Jin-Ho Lim3. 1Maria Fertility Hospital, Seoul, Korea,, common infertility phenotype in North Africa population.
2
Chonbuk National University, Jeonju, Korea, 3Fertility Research Center, Maria
Medical Foundation, Seoul, Korea. Material and Methods: Case report -Our observation is about an infertile 35-
year-old man unable to conceive despite 2 years of unprotected intercourse.
Introduction: The major benefits of in vitro maturation (IVM) include This patient stemming from a consanguine marriage and presenting a
avoidance of the risk of ovarian hyperstimulation syndrome (OHSS), reduced familial history that revealed multiple cases of infertility, perinatal or low age
cost, and less complicated treatment for patients. This study reports the deaths. Repeated semen analysis revealed a severe oligoasthenospermia
clinical outcome for 8 years of our IVM-ET program. and necrospermia with sperm count < 1 Million spz /ml, total motility <5%,
Materials and Methods: This study was performed retrospectively. A total of vitality< 20%. Cytomorphological analysis using shorr coloration revealed a
717 IVM cycles were analyzed from May 2000 to December 2007. Oocytes severe monomorphic teratozoospermia with 100% of abnormal
were collected between day 7 and 13 of the menstrual cycle. The ovum pick- macrocephalic sperm heads. Moreover, 70% of spermatozoa carried
up day was estimated from over-all cycle length and endometrial thickness. multiple flagella. The midpiece was enlarged with cytoplasmic droplets in
The patients were administered 10,000 IU HCG 36 h prior to oocyte 52% of spermatozoa. The multiple anomalies index (MAI) was 3.9 (normal
collection. A transvaginal ultrasound machine with a 19-gauge aspiration value = 1.6), reflecting the high incidence of spermatozoal morphological
needle was used to aspirate follicles. Follicular aspirates were filtered using abnormalities in this patient. Chromosomal analysis revealed a normal
70-mm mesh, and then the oocytes were isolated under a karyotype (46,XY). A genetic factor was strongly suspected and a molecular
stereomicroscope. Immature oocytes were cultured in YS maturation genotyping was assessed by a genome-wide microsatellite scan.
medium. Fertilization of matured oocytes was induced by ICSI. 2PN zygotes Results: A homozygous mutation (c.144delC) in the Aurora Kinase C
were co-cultured with cumulus cells in 20 of YS culture medium. Good (AURKC) gene was identified with a single nucleotide deletion in the AURKC
quality embryos were transferred on day 4 or 6 after oocyte retrieval. coding sequence. This founder mutation results in premature termination of
Results: A total of 11,665 oocytes were retrieved from 717cycles (mean age translation, yielding a truncated protein that lacks the kinase domain.
= 32.0 ± 3.5 years). The maturation rate was 70.9% (8,269/11,665) and Conclusions: Recent studies demonstrated that a homozygous mutation
fertilization rate was 78.3% (6,478/8,269). Following transfer of embryos, (c.144delC) in the Aurora Kinase C (AURKC) gene led to a meiotic division
clinical pregnancy was achieved in 223 cycles (31.1%, 223/717) with an deficiency and the production of large-headed multi-flagellar spermatozoa, a
implantation rate of 10.0% (330/3,289). primary infertility phenotype mainly observed in the Maghreb. Aurora Kinase
Conclusions: An acceptable pregnancy rate can be achieved through an IVM- C is a protein expressed in testis and implicated in the meiotic segregation
ET program. These results suggest that in vitro maturation is a clinically during spermatogenesis. This molecular defect leads to the genesis of
useful method for polycystic ovarian syndrome patients who have risks polyploid spermatozoa with abnormal cytogenetic content. The treatment of
associated with ovarian stimulation. these cases of infertility using ICSI would be unsuccessful and have a high
genetic risk.
April 19-22, 2009 96

1 group were frozen without any exposure to CLC or MBCD and in control 2
P.090 – EVALUATION OF RIF PATIENTS WITH RESPECT TO TYPE OF SPERM (vehicle), sperms were incubated with 4mM MBCD.Then the post-thaw
SELECTED FOR IMSI sperms were evaluated.
Semra Mılık, Zafer Atayurt, Sevil,Unal, Hakan Yelke, Sebnem Yazıcı, Ferhat Results: The values of the intact acrosome, motility and fertilizing ability
Cengiz, Semra Kahraman. Memorial Hospital IVF Center, Istanbul, Turkey. increased significantly with concentration of CLC compared to controls and
MBCD experimental groups (P<0.05).
Objective: To evaluate the effect of the sperm grade, selected at high
magnification according to their morphology and existence of nuclear Conclusion: These results indicate that cryosurvival of C57BL/6 mouse
vacuoles, on subsequent embryo development and pregnancy rates. spermatozoa is enhanced by exposure to MBCD which loaded with
cholesterol (CLC) before freezing.
Design: Comparative prospective study
Materials and Methods: 81 couples having at least two previous unsuccesful
trials were included in the study. Spermatozoa used for IMSI were classified P.093 – THE PROTECTIVE EFFECTS OF CARROT SEEDS EXTRACT ON
according to morphology of the head and existence and size of the nuclear SPERMATOGENESIS AND CAUDA EPIDIDYMAL SPERM RESERVES IN
vacuoles as Grade I:normal head morphology and no vacuoles, Grade II: GENTAMICIN TREATED RATS
:normal head morphology and one small vacuole (4% of head surface), Mohammad Nouri, Arash Khaki, Fatemeh Fathi Azar, Mohammad-Reza Rashidi.
Grade III:normal head morphology and more than one small vacuole, Grade Tabriz University of Medical Sciences, Tabriz, East Azarbaijan, Iran.
IV:abnormal head morphology and large vacuoles, a system previously Background: Carrot (Daucus carota L.) is known to possess antifertility
described by Vanderzwalmenn et al. Spermatozoa having defects on properties in female. However, according to Iranian traditional medicine, the
neck,mid-piece and tail were not selected for microinjection, regardless of effect of carrot on fertility is gender dependent and this food can increase
head morphology. Sperm selection was made by Zeiss Axioobserver A1 the potency in men. The aim of this study was to investigate the influence of
inverted light microscope, equipped with DIC optic system and Narishige carrot seed extract (CSE) on spermatogenesis, number and motility of
micromanipulators, at a total of 8050X magnification on monitor. Patients sperms in cauda epididyme in male rats.
were grouped in three according to the grade of the sperms used as, Grade I
for 36 of the cases, Grade II for 31 and GradeIII or IV for 14. There were no Materials and Methods: Forty adult male rats were randomly divided into 5
differences between groups with respect to mean age of women (33,1±5,5, groups: normal group, groups receiving low- and high doses of CSE,
31,5±5,5 and 31,5±5,2, respectively) animals that received high-dose of CSE with gentamicin, and gentamicin
treated group. After 4 weeks treatment, serum samples were obtained for
Results: 200,208 and 61 oocytes were injected for Groups 1,2 and 3 the sex hormone analysis. Under anesthesia, testis, cauda epididymides and
respectively. Fertilization rates were similar between groups 1 and 2 (79.5% sperm ducts were dissected and sperm count, motility and cauda
vs. %80.8%) but lower for the third group (70,9%). We have found also a epididymis sperm reserves were determined. Histopathological changes of
similar relationship between groups for the top quality embryos (more than testis were also studied to assess spermatogenesis. Data analysis was
6 blastomers,evenly sized and fragmentation lower than 10%) at day 3 performed using one-way ANOVA followed by Tukey HSD tests.
(45,2%,46,4% and 34,5%) and clinical pregnancy rates (40,9%,41,9% and
Results: Administration of CSE caused a significant increase in the cauda
epididymal sperm reserves (CESR) compared with the control (28.2 ± 1.8
35,7) as Groups 1 and 2 are similar and Group 3 is lower.
Conclusion: Our results have shown and also contribute to the previous vs. 45.1 ± 2.0, ↔106). The extract could also protect testis from the
findings of similar studies that the normalcy of sperm selected for IMSI gentamicin-induced necrosis. The CSE administration caused about 3.5-
affects the outcome positively. So it is important to search for the best times increase in the LH levels of the rats even in spite of receiving 5
sperm when it is possible. But since there is no significant difference mg/kg/day gentamicin with no significant effect on FSH levels. The
between the results of Grade I and II sperms, spending too much time to testosterone concentrations in the group received 400 mg/kg CSE were 30%
find the best sperm should be avoided, taking into account the possible and 83% higher than its levels in the control and the gentamicin treated
negative effect of incubation in PVP. group, respectively.
Conclusion: CSE is able to overcome reproductive toxicity of gentamicin and
P.091 – COMPARISON THE EFFECTS OF CHOLESTEROL AND METHYL-BETA- induces spermatogenesis probably mainly through the elevation of
CYCLODEXTRIN ON CRYOSURVIVAL OF C57BL/6 MOUSE testosterone levels. It appears that this extract has opposite effects on male
SPERMATOZOA and female reproductive systems.
Shabnam Movassaghi1, Ghasem Saki2, Fatemeh Javadnia2, Marzieh Panahi2
Mahmoud Mahmoudi3; 1Department of Anatomy, School of Medicine, Islamic Azad P.095 – THE HISTOPATHOLOGIC EFFECTS OF ELECTROMAGNETIC FIELD ON
University-Tehran Medical Branch, 2Department of Anatomy, physiology research PREIMPLANTATION PHASE OF MOUSE OVARY
center, School of Medicine, Jondishapour University of Medical Sciences, Ahwaz,
Iran, 3Department of Biostatistics, School of Medicine, Tehran University of Farzad Rajaei, Fatemeh Sabbagh Ziarani. Qazvin University of Medical
Medical Sciences; Tehran, Iran. Sciences, Qazvin, Iran.
Introduction: Cryopreservation induces damage to mouse sperm especially Introduction: Life on earth has evolved in a sea of natural electromagnetic
C57BL/6 mouse strain and results in a loss of motile and viable cells. Part of fields (EMFs). Human data reviewed concern the potential reproductive
this damage occurs due to membrane alterations induced by the membrane effects (mainly, spontaneous abortions, low birth weight, and congenital
changing from the fluid to the gel-state as the temperature decreased. malformations) of exposure to various sources of EMFs.
Methyl-beta-cyclodextrin (MBCD), which leads to the stimulation of Materials and methods: 80 female mice were divided in to 2 groups. Control
cholesterol efflux from the cell membrane, is capable to improving group was not exposed to EMF and case group was exposed to 4 hours per
cryosurvival of frozen/thawed sperm in some mammalian species. But day, 6 days a week for 2 weeks to 50 Hz & 0.5 mT EMF. Female mice in both
another observations suggest that adding cholesterol to sperm plasma groups on 8th day of exposing were superovulated and mated over the
membrane may increase the membrane fluidity during cryopreservation. night. Next morning females with a vaginal plug were identified as pregnant
MBCD and cholesterol-loaded-cyclodextrin (CLC) were examined for their mice; at the time of implantation, the pregnant mice were killed and
abilities to increase the cryosurvival of C57BL/6 mouse sperm. The blastocysts were subsequently obtained from these mice by flushing the
intactness of acrosome, motility and fertilizing ability of frozen/thawed uterus horns. The samples of ovaries in all groups were taken and were
spermatozoa were used to monitor cryosurvival. processed for light microscopic studies. The data have been compared
using statistical methods (SPSS, t test and P<0.05).
Methods: This experimental study was performed in Cell Culture Laboratory
of Ahwaz Jondishapour University of Medical Sciences during autumn and Results: Results showed that the mean number of pregnant mice decreased
winter 2007. male mice were randomly divided in six groups: In in EMF group) 50%) compared to the control group) 67.5 %( but the
experimental groups spermatozoa were exposed to different concentrations difference between them was not significant. The mean number of fetuses
of CLC or MBCD and subsequently cryopreserved. Spermatozoa in control per pregnancy was (9±4.8) in control group and (5.5±5.7) in experimental
97 Geneva
group and statistical analysis were showed significant decrease between

mean of 2 groups (P<0.03). The analysis of the size of monolayer primary
follicle in EMF exposed groups did not show significant decrease in compare
to control group (107.22±13.39, 105.86± 15.63 and P>0.810). Although the
total number of follicles, number of monolayer primary follicle & corpus
luteum, increase in compare to control group respectively but there was no
significant differences between them.
Conclusion: The findings indicated that the EMF in short period of exposure
has negative effects on female mice fertility but histological studies show no
negative effects on ovaries.
P.096 – RELATIONSHIP BETWEEN SERUM ESTRADIOL LEVEL AND OOCYTE

AND EMBRYO QUALITY FROM ICSI TRIALS
S. Masadnah, M.A Danfour, O.A.,Elsrait, Mohamed S. Elmahaishi. Misurata
Infertility Centre, Misurata, Libya.
Oocyte quality affects early embryonic survival, the establishment and
maintenance of pregnancy, fetal development, and even adult disease.
Quality, or developmental competence, is acquired during folliculogenesis as
the oocyte grows, and during the period of oocyte maturation. Assisted
reproductive technologies involving ovarian hyperstimulation, perturb this
process and result in oocytes with reduced quality. The aim of the study
was to investigate the influence of serum oestradiol level in women who
underwent intracytoplasmic Sperm Injection (ICSI) program oocyte and
embryo production quality.
Study Design: Retrospective, comparative.
Material and Methods: 600 patients were included. There were divided in
three groups depending on the serum oestradiol level: Group A:(<300
pg/mL, group B: 300-500 pg/mL and group C: > 500 pg/mL). Therapeutic
protocol. All women were ≤42 years of age (average age 33; range 25-42.
Ovarian stimulation was performed with gonadotrophins (Puregon &
Menogon or Minipure) (short protocol) following down regulation of the
pituitary with a gonadotrophin releasing hormone agonist (Gonopeptyl).
When the leading follicle reached a mean diameter of 18-20 mm and serum
oestradiol appeared adequate, human chorionic gonadotrophin (hCG,
10,000 IU) were injected.
Statistical analysis: (SPSS 11) with one way ANOVA and person correlation
was used to analyze the patients age, number of metaphase II oocytes
retrieval, oocyte quality, oocyte maturation, oocytes fertilized, embryo
cleavage and number, quality of transferred embryos and and serum βhCG
among three groups.
Results: There were statistically significant correlation between oocyte
number, serum βhCG among three groups and estradiol level, however there
were no correlation between serum oestradiol level and oocyte quality,
oocyte maturation, oocytes fertilized, embryo cleavage and number, quality
of transferred embryos among three groups.
Conclusion: These retrospective data demonstrated that oestradiol serum
level do not affect the quality of oocyte, potential fertilization and embryo
developmental competence, however it may affect implantation level.
Therefore serum oestradiol level may predict ICSI outcome and considered
as prognostic value.
April 19-22, 2009 98

EXHIBIT DIRECTORY
99 Geneva
FLOOR PLAN
April 19-22, 2009 100

EXHIBIT DIRECTORY
Exhibit Hours
Sunday, April 19 18:00-20:00 Official Welcome Reception

Monday, April 20 08:30-18:00
Tuesday, April 21 08:30-18:00
Wednesday, April 22 08:30-18:00
List of Exhibitors in Alphabetical Order
Co m p a n y Na m e B o o t h Nu m b e r
CCD Lab 19
Cryo Bio Systems 11
Eppendorf AG 17
European Sperm Bank 14
Ferring Pharmaceuticals 32-33
GE Healthcare 38
Gynemed GmbH 24
IBSA Laboratories 15-16-27-28
Irvine Scientific 10
IUL Instruments/Ruskinn Life Sciences 8-9
IVF Online 18
ISIVF & IVM Behind Registration
Jaypee Brothers Medical Publishing 12
Karl Storz GmbH & Co KG 36-37
McGill Symposium Behind Registration
MediCult 31
Merck-Serono 42-43-44
MTG - Medical Technology Vertriebs-GmbH 30
Nanopoint Imaging 26
Nikon 39
Olympus Life Science Europa GmbH 34
Research Instruments Ltd. 6
Schering-Plough 40-41
Smiths Medical 25
Sparmed APS 5
Unisense 35
Vitrolife 7
Wisepress
101 Geneva
EXHIBITOR DETAILS
CCD INTERNATIONAL EPPENDORF AG
BOOTH: 19 BOOTH: 17
Mrs GIBAULT Catherine Dr. Heide Niesalla
48, rue des Petites Ecuries Barkhausenweg 1
75010 Paris 22339 Hamburg
France Germany
Telephone: +33 1 34 44 15 15 Telephone: +49 40 53801 - 0
Fax: +33 1 30 72 22 08 Fax: +49 40 53801 - 556
E-Mail: cgibault@ccdlab.com E-mail: eppendorf@eppendorf.com
Website: www.ccdlab.com Website: www.eppendorf.com
C.C.D. International has a long-standing tradition of developing medical Eppendorf is a biotechnology company that develops, manufactures and
devices for IVF and general OB/GYN practice. Our core competences include distributes systems comprising instruments, consumables and reagents for
extensive experience in the design, development and manufacturing of use in laboratories worldwide. The product range includes systems for cell
catheters and needles with state of the art, medical grade materials. Our manipulation and microinjection, liquid handling, centrifugation as well as
scientists and engineers bring extensive skills and broad experience toward complete equipment for DNA amplification and biochips.
delivering the utmost in quality, design performance, reliability and value.
Many C.C.D. devices are worldwide gold-standards such as Frydman® EUROPEAN SPERM BANK
catheters, Echogyn® Embryoview®, Pipelle® by Dr Cornier, Pipelle® Mark
II, H Pipelle®, S.I.S. Rudigoz catheter. Our new release, the ART of CCD® BOOTH: 14
line for oocyte retrieval, has been designed to meet the most demanding Dr. Peter Bower, Ph.D.
needs of experts in assisted reproductive techniques. Falkoner Alle 63, 2
CRYO BIO SYSTEM DK-2000 Frederiksberg, Copenhagen
Denmark
BOOTH: 11
Mrs. Béatrice Ledos Telephone: +45 38343600
29 rue Tronchet Fax: +45 38343646
75008 Paris E-mail: bower@europeanspermbank.com
France Website: www.europeanspermbank.com
The European Sperm Bank is among the leading sperm banks in Europe. The
Telephone: +33 (0)149 240 505 European Sperm Bank offers patients a choice of highly screened donor
Fax: +33 (0)149 240 501 sperm and to have it shipped to their clinic. We offer both Anonymous and
E-mail: contact@cryobiosystem-imv.com Open Identity donors, and all of our donors have IUI-ready samples.
Website: www.cryobiosystem-imv.com
FERRING PHARMACEUTICALS
Cryo Bio System, the expert in cryopreservation with its CBS™ High
Security straw concept, is now worldwide recognized for its HSV High BOOTH: 32-33
Security Vitrification Kit, a reliable, fast and secure device for oocyte and Ms. Elisabeth Weis
embryo vitrification. Cryo Bio System also manufactures IUI/ET catheters and Chemin de la Vergognausaz 50
brand-new equipment for the ART and Biobanking markets.
CH-1162 Saint-Prex
Switzerland
Telephone: +41 58 301 00 00
Fax: +41 58 3010371
E-mail: info@Ferring.com
Website: www.ferring.com
Ferring is a Swiss-based research driven, specialty biopharmaceutical group
active in global markets. The company identifies, develops and markets
innovative products in the areas of fertility, gynecology, endocrinology,
gastroenterology and urology. In recent years Ferring has expanded beyond
its traditional European base and has offices in over 40 countries. To learn
more about Ferring or its products, please visit www.ferring.com and come
to the FERRING booth.
April 19-22, 2009 102

EXHIBITOR DETAILS
GE HEALTHCARE IRVINE SCIENTIFIC
BOOTH: 38 BOOTH: 10
Mrs. Martina Emde Ms. Kiersten Carlin
Beethovenstrasse 239 2511 Daimler Street
D-42665 Solingen Santa Ana, CA, 92705
Germany USA
Telephone: +49 212 2802258 Telephone: +1 949 261-7800
Fax: +49 212 2802439 Fax: +1 949 261-6522
E-mail: martina.emde@med.ge.com Website: www.irvinesci.com
Website: www.med.ge.com Irvine Scientific is the world leader in providing a complete line of media and
GE is dedicated to helping you transform healthcare delivery by driving supplies for Assisted Reproductive Technologies. The success of our
critical breakthroughs in biology and technology. Our expertise in medical customers is a direct result of our commitment to quality, consistency and
imaging, medical diagnostics, patient monitoring systems and drug reliability. Irvine Scientific is the exclusive U.S. distributor for Wallace
discovery is enabling healthcare professionals around the world discover needles and catheters.
new ways to predict, diagnose and treat disease earlier. For additional
information visit: www.gehealthcare.com IUL INSTRUMENTS GMBH
BOOTH: 9
GYNEMED MEDIZINPRODUKTE GMBH & CO. KG Königswinterer Str. 409a
BOOTH: 24 D-53639 Königswinter
Mrs. Andrea Andresen Germany
Radeberstr. 21
Telephone: +49-02223-9192-26
D-23738 Lensahn
Website: www.iul-instruments.de
Germany
IUL Instruments GmbH is a leading provider of high quality laboratory
Telephone: +49/4363/903290 equipment to life science customers in Germany, Switzerland and Austria.
Fax: +49/4363/9032919 We pride ourselves on our scientific expertise in microbiology, cell biology
and biotechnology and on our excellent support of customers. Ruskinn and
E-Mail: aan@gynemed.de IUL Instruments will be happy to demonstrate the Ac-tive™ IVF System for
Website: www.gynemed.de ART laboratories at booths 8 & 9.
Gynemed presents the GM501 media line, developed for handling and
culture of human germ cells and embryos with biggest success. The GM501 RUSKINN LIFE SCIENCES LTD.
line offers a complete range of media needed in the IVF-lab. The media meet (IN COOPERATION WITH IUL INSTRUMENTS GmbH)
the latest scientific cognitions and are established as standard in many labs
worldwide. BOOTH: 8
Website: www.ruskinn.com
IBSA INSTITUT BIOCHIMIQUE SA Ruskinn is one of the world’s leading suppliers of modified atmosphere
BOOTH: 15-16-27-28 workstations. In July 2008, Ruskinn introduced the Ac-tive™ IVF System, a
Mrs. Elisabetta Artioli, Pharm. D. totally enclosed ART Workstation where all manipulations, embryo culture
and microscopic examinations can take place. Ac-tive™ significantly reduces
Via del Piano gamete and embryo stress, thus increasing pregnancy rates.
CH- 6915 Pambio Noranco, Lugano
Switzerland IVFONLINE
Telephone: +41-58-3601624 Booth: 18
Fax: +41-58-3601647 Telephone: +1 519-826-5800
E-mail: elisabetta.artioli@ibsa.ch Fax: +1 519-826-6947
Website: www.ibsa.ch Website: www.IVFonline.com
IBSA is an international pharmaceutical company headquartered in Lugano, LifeGlobal®: Increased worldwide success of global media; the only
Switzerland, delivering tailored therapeutic solutions for follicular stimulation scientifically-based and clinically-proven single culture medium. SunIVF®:
and luteal support. IBSA’s whole in-house manufacturing cycle provides New dishes for larger volume culture. GPS™, Corral®, Universal - the safest
highly purified and highly glycosylated, human-derived gonadotrophins such dishes for IVF Coda®: New ECO Tower; continued success of Coda®
as hFSH (Fostimon), hMG (Merional), hCG (Choriomon). Other company's Inlines - good standard practice. Free Fertility Magazine.
franchises include osteoarthritis, pain-management, dermatology and thyroid
diseases.
103 Geneva
EXHIBITOR DETAILS
JAYPEE BROTHERS MEDICAL MEDICULT
PUBLISHERS (P) LTD. BOOTH: 31
BOOTH: 12 Monsieur Denis AZRA, MediCult France
Mr. J.P. Vij 1 rue des Vergers
4838/24, Ansari Road, Daryaganj F-69760 Limonest
New Delhi 110 002 France
India Telephone: + 33 472 564 800
Telephone: +91-11-43574357 Fax: + 33 472 564 801
Fax: +91-11-43574314 E-mail: medicult-france@wanadoo.fr
E-mail: jaypee@jaypeebrothers.com MediCult is a leader in delivering leading innovative ART solutions to the
Website: www.jaypeebrothers.com benefit of families. Through innovation and product advancements, MediCult
has acquired in 2007 Humagen and in 2008 MidAtlantic Diagnostics;
Jaypee Brothers is a medical publisher who publishes maximum obgyn MediCult aims to help the # 1 dream of every infertile couple come true.
books. So far they have published about 200 titles exclusively in Obstetrics
& Gynecology. They are a very high quality book publisher and the prices
are within the reach of the doctors and that is the secret of the popularity of
MERCK SERONO
Jaypee’s Obgyn books in the world. BOOTH: 42-43-44
Ms. Mel Lewis
KARL STORZ GMBH & CO. KG Telephone: +41 22 414 4778
BOOTH: 35-36 Fax: +41 22 414 3003
Ms. Sigrid Lanzillotti E-mail: mel.lewis@merckserono.net
Mittelstrasse 8 Website: www.merckserono.com or www.merck.de
D-72538 Tuttlingen
Merck Serono is the division for innovative prescription pharmaceuticals of
Germany Merck, a global pharmaceutical and chemical group. Headquartered in
Geneva, Switzerland, Merck Serono discovers, develops, manufactures and
Telephone: +49 7461 7080 markets innovative small molecules and biopharmaceuticals to help patients
Fax: +49 7461 708 105 with unmet medical needs. Its North American business operates in the
E-mail: info@karlstorz.de United States and Canada as EMD Serono.
Website: www.karlstorz.de Merck Serono has leading brands serving patients with cancer (Erbitux®),
multiple sclerosis (Rebif®), infertility (Gonal-f®), endocrine and metabolic
KARL STORZ is a renowned manufacturer that is well established in all fields disorders (Saizen®, Serostim®, Glucophage®, Concor®, Euthyrox®,
of endoscopy. The still family held Company has grown to one with a Kuvan®).
With an annual R&D expenditure of around €1bn, Merck Serono is
worldwide presence and 4000 employees. KARL STORZ offers a range of
both rigid and flexible endoscopes for a broad variety of applications.
committed to growing its business in specialist-focused therapeutic areas
McGILL SYMPOSIUM, IVM & EGG FREEZING including neurodegenerative diseases, oncology, fertility and endocrinology,
as well as new areas potentially arising out of research and development in
WORKSHOPS autoimmune and inflammatory diseases.
Telephone: +1-514-843-1729
MTG MEDICAL TECHNOLOGY VERTRIEBS-GMBH
Fax: +1-514-843-1476
BOOTH: 30
E-mail: info@mcgillsymposium.com
Opalstrasse 32
Website: www.mcgillsymposium.com
D-84032 Altdorf
Learn first-hand how to incorporate IVM within your practice. Discover the
techniques of immature oocyte retrieval and identification. Acquire the latest
Germany
knowledge of Cryobiology techniques performing vitrification of mouse Telephone: +49 871 97519-0
oocytes using the McGill Cryoleaf. Participants will be able to implement the
learned techniques into their programs upon completion of this course. Fax: +49 871 97519-70
Website: www.mtg-de.com
MTG – Your expert in advanced A.R.T. technology. We will be displaying pH
OnlineTM for continuous pH recording from the incubator as well as the
major components of the integrated OCTAX platform: OCTAX Laser Shot™,
OCTAX polarAIDETM for comprehensive spindle and zona analysis, OCTAX
cytoScreenTM for real-time zoomed sperm analysis (“dry IMSI”).
April 19-22, 2009 104

EXHIBITOR DETAILS
NANOPOINT, INC. RESEARCH INSTRUMENTS LTD.
BOOTH: 26 BOOTH: 6
Ms. Cathy Owen Mr. Justin Retallack
900 Fort Street Mall, Suite A20 Bickland Industrial Park
Honolulu, Hawaii, 96813 Falmouth, Cornwall, TR11 4TA
USA United Kingdom
Telephone: +1-808-457-1145 Telephone: +44 (0) 1326 372753
Fax: +1-808-537-4245 Fax: +44 (0) 1326 378783
E-mail: info@nanopointimaging.com E-mail: justin@research-instruments.com
Website: www.nanopointimaging.com Website: www.research-instruments.com
Nanopoint, Inc. is a privately-held biotechnology company that is advancing Research Instruments will be displaying our Saturn Laser system, Integra Ti
the study and treatment of diseases with its live cell imaging solutions. micromanipulation system, new EZ-Strip denudation system and a range of
Nanopoint's cellTRAY® Imaging Systems with integrated microfluidics have pipettes. Also on show is a complete range of environmental monitors, as
broad applications to life science research, drug discovery, and assisted well as the IVF Witness automated gamete tracking system and IVF Tracker
reproductive technology. for tracking consumables.
NIKON INSTRUMENTS AG SCHERING-PLOUGH

BOOTH: 39 BOOTH: 40-41
Mr. Lukas Jufer Mr. Ralph Zürcher
Im Hanselmaa 10 ESSEX Chemie AG
CH-8132 Egg/ZH Weystrasse 20
Switzerland CH-6006 Lucerne
Telephone: +41 43 277 28 67 Switzerland
Fax: +41 43 277 28 61 Telephone: +41 79 358 03 18
E-mail: lukas.jufer@nikon.ch Fax: +41 41 368 49 36
Website: www.nikon.ch E-Mail: ralph.zuercher@spcorp.com
From being instrumental in the first IVF births, to developing latest
Website: www.schering-plough.com/
technologies, two core values are at the heart of Nikon’s IVF offerings: Essex Chemie AG, based in Lucerne, is the Swiss country operation of
commitment to optical excellence; and an ethos of developing embryo- Schering-Plough Corporation (S-P), based in Kenilworth, New Jersey, USA.
friendly workflows. Please come and see how Nikon’s latest developments In Switzerland Essex Chemie AG markets a broad range of innovative human
could revolutionise embryology and remove stress: from the embryo’s and prescription products with the emphasis on allergology, anesthesia, CNS,
from the embryologist! dermatology, fertility, gynecology, hepatology, cardiovascular, immunology,
mycology and oncology. Through its own biopharmaceutical research and
OLYMPUS LIFE SCIENCE EUROPA GMBH collaborations with partners, S-P creates therapies that help save and
improve lives around the world. The vision of the company is to “Earn Trust,
BOOTH: 34 Every Day” with the doctors, patients, customers and other stakeholders
Dr. Friederike Lehmann served by its colleagues around the world.
Wendenstrasse 14-18
D-20097 Hamburg
Germany
Telephone: +49 40 23773 5406
Fax: +49 40 23773 4647
E-mail: friederike.lehmann@olympus-europa.com
Website: www.microscopy.olympus.eu/microscopes/
Total clarity; total comfort Olympus is displaying the advanced IX71 inverted
microscope system for easy and comfortable cell imaging. With a unique
two-tiered multi-port design for maximum flexibility, the IX71 gives brighter
images and modes for fluorescence, DIC, phase contrast and Olympus Relief
Contrast. Also on display: micromanipulators and microinjectors from
Eppendorf.
105 Geneva
EXHIBITOR DETAILS
SMITHS MEDICAL INTERNATIONAL LTD. VITROLIFE SWEDEN AB
BOOTH: 25 BOOTH: 7
Miss Sarah Fry Ms. Sara Aurelius
Portex House, Military Road P.O. Box 9080
Hythe, Kent, CT21 6JL SE-400 92 Göteborg
United Kingdom Sweden
Telephone: + 44 1303 236936 Telephone: +46 31 721 80 00
Fax: + 44 1303 236777 Fax: +46 31 721 80 90
E-Mail: sarah.fry@smiths-medical.com Website: http://www.vitrolife.com
Website: www.smiths-medical.com We have over 25 years of experience in developing high quality products for
Smiths Medical worked with the pioneers in IVF in the design and ART covering all steps of an IVF-treatment. Welcome to our booth to learn
manufacture of the Wallace Classic Embryo Replacement Catheter and is more about the latest news in our product portfolio. Get the latest data on
associated with some of the highest pregnancy rates in the world. Smiths the G5 Series™ and take a closer look at - Swemed Sense™ - a new needle
Medical also manufactures a Single and Dual Lumen Oocyte Recovery designed to minimize tissue damage, bleeding and patient discomfort.
Needle range.
WISEPRESS ONLINE BOOKSHOP
SPARMED The Old Lamp Works, 25 High Path, Merton Abbey
BOOTH: 5 London, SW19 2JL
Mr. Onur Ozturk United Kingdom
Farum Gydevej 89 Telephone: +44 20 8715 1812
DK-3520 Farum Fax: +44 20 8715 1722
Denmark E-mail: bookshop@wisepress.com
Telephone: +45 39 40 25 03 Website: www.wisepress.com
Fax: +45 39 40 25 64 Wisepress.com, Europe’s leading conference bookseller, has a complete
E-Mail: info@sparmed.dk range of books and journals relevant to the themes of the meeting. Books
can be purchased at the stand or, if you would rather not carry them, posted
Website: www.sparmed.dk to you – Wisepress will deliver worldwide. In addition to attending 250
Sparmed is a Danish company which is worldwide exclusive distributer for conferences per year, Wisepress has a comprehensive medical and scientific
IVFtech’s Oosafe® brand products such as Oosafe® Filter and Oosafe® Air bookshop online with great offers, some up to 40% off the publisher list
Cleaner. We aim to bring new products to ivf market cooperating with prices.
IVFtech which is the manufacturer of ivf workstations, anti-vibration tables,
mini incubators and heating systems for ivf laboratory use. Sparmed is a
turnkey IVF laboratory projects and clean air solutions expert.
UNISENSE FERTILITECH A/S

BOOTH: 35
Ms. Francesca Bahr
Brendstrupgaardsvej 21F
DK-8410 Aarhus
Denmark
Telephone: +45 89449525
Fax: +45 89449549
E-mail: fab@unisense.com
Website: www.unisense.com
Unisense Fertilitech A/S presents the EmbryoScope™ Advanced Embryo
Incubator, which allows time-lapse image acquisition of up to 72 developing
embryos while maintaining a safe and fully controlled incubator
environment. The EmbryoScope™ Incubator will facilitate selection of the
best viable embryo, while minimizing disruptions to embryo development.
April 19-22, 2009 106

PRESENTER INDEX
Abdallah, Michel Abou 401.4 Farimani, M. P.082
Abdollahi-Fard, Seddigheh P.001 Feki, Anis 102.3, 402.1
Abe, Hiroyuki P.002 Feldman, Lisa 105.2
Abir, Ronit 306.2 Fenichel, Patrick 408.3
Aboulghar, Mohamed 404.4 Ficicioglu, Cem P.031, P.032
Ahuja, Kamal 404.2 Fındıklı, Necati 210.2
Ajina, Mounir P.006 Frydman, Nelly 101.3, 209.2
Akbarpou, Mahzad P.008 Frydman, René 101.1, 205.4, 311.2
Akdogan, Aysin P.009 Fulka Jr., Josef 208.1
Albertini, David 208.3 Genazzani, Andrea Riccardo 309.6
Alipour, Hiva P.079 Germond, Marc 301.4
Alizadeh, Leila P.011 Ghahremani, Manda 312.7
Ambrosetti, Alexandra 206.6, P.012 Gianaroli, Luca 311.3
Amiri, Iraj P.013 Gilbert, Lucy 305.1
Aniya, Tomoka P.014 Gneist, Nadja 211.4
Aono, Fumihito P.015 Göker, Ege 210.3
Arabzadeh, S. P.039 Gomel, Victor 100.3, 403.4
Argyrou, Marina P.016 Gougeon, Alain 300.2
Asherkaci, Hussian 410.1 Gülekli, Bülent 310.1
Azadbakht, Mehri P.017, P.018 Gürgan, Timur 104.2, 204.2
Bahar, L. P.019 Gutnisky, Cynthia P.033
Bahçeci, M. 210.6 Haloob, A. Rahim 312.4
Balawi, M. P.020 Hammadeh, M.E. 207.6, P.034
Barak, Yona 304.1 Hanck, Beverly 409.3
Belaisch-Allart, Joelle 301.3 Handyside, Alan 101.2, 200.1, 203.2, 203.3
Belloc, Stéphanie 309.3 Haruki, Atsushi P.035
Ben-Yosef, Dalit 402.2 Hassani Bafrani, Hassan P.036, P.037
Benkhalifa, Moncef 104.3, 207.2, 210.1, 406.1, P.021 Holzer, Hananel 405.2
Bernard, Artur 303.1 Hovatta, Outi 102.1, 105.4, 209.1, 400.1
Bersinger, N. 206.5 Ida, Mamoru P.040
Bischof, Paul 201.3 Imthurn, Bruno 209.3
Boivin, Jacky 409.5 Ishizuka, Bunpei 205.1
Bouchard, Philippe 205.2 Jayakrishnan, Kay 211.6, P.044
Buckett, William 209.4 Jones, Keith T. 405.1
Campbell, Keith 400.2 Josso, Nathalie 408.2
Cha, Jeong-Ho P.081 Kaluarachchi, Athula 407.1, 407.2
Chae, Soo Jin P.050 Kamrava, Michael 312.3
Chian, Ri-Cheng 302.2 Kang, Hee Jung P.042, P.043
Chillik, Claudio 404.3 Kato, Keiichi 303.2
Cho, Hwang-Yun P.023 Kato, Osamu 401.3
Choi, Hye Won P.024 Kazaryan, Liya P.045
Dal Canto, Mariabeatrice 405.3 Keck, Gudrun P.046
Danfour, Mohamed 211.1 Keskes, L. P.087
de Geyter, Christian 401.1 Kossowska-Tomaszczuk, K. 206.3
Demirol, Aygül 204.5, 210.4, 302.3 Kostyuchek, Irina 211.2, P.048
Dogan, Muammer P.071 Kovacs, Peter 307.1
Dokuzeylul, Nur P.025, P.026 Kuwayama, Masashige 307.3
Donnez, Jacques 100.1, 300.1, 305.3 Kyono, Koichi 308.3
Dor, Jehoshua 302.1 Ledger, William 405.4
Dorfmann, Andrew 410.6 Lee, Shaw Ni Amy P.049
Dreyfus, Jean-Michel 407.7 Leong, Milton 208.5
Dubuisson, Jean-Bernard 100.2, 403.2 Lessey, Bruce 201.2
El-Garem, Yehia P.027 Leung, Peter C.K. 205.3
Elmahaishi, Mohamed S. P.096 Lim, Jin Ho 200.2
Elsraite, Omar P.028 Lindenberg, Svend 310.4
Es-slami, Samira P.029 López, Gemma P.051
Fabbri, Raffaella 410.3 Macas, M. 206.1
Fadini, Rubens 208.4 Mahmoudi, Reza P.052
Fanchin, Renato 408.4 Mansour, Ragaa 406.2
Fancsovits, P. P.030 Marinakis, Gerasimos 211.5
107 Geneva
PRESENTER INDEX
Massarwa, Abir 407.6 Tepekoy, Filiz 309.4
Mencaglia, Luca 403.3 Tijani, H. P.010
Mengistu, Meserat 105.3 Tirelli, Alessandra P.070
Mettler, Liselotte 204.4, 407.5 Tomazevic, Tomaz 407.3
Mikkelsen, Ann Lis 310.2 Tuong, H.M. P.038
Miyaki, Yukiko P.054 Tur-Kaspa, Ilan 201.4, 301.2, 103.2
Mılık, Semra P.090 Ueno, Keiko P.072
Morimoto, Yoshiharu 304.3, 306.3 Ulug, U. 210.7
Movahedin, M. P.055 Urner, F. 206.2
Movassaghi, Shabnam P.091 Utsunomiya, Takafumi 307.2, P.073
Mueller, Michael 100.4 van der Poel, Sheryl 105.1, 409.1
Munk, Mette 309.2 van Herendael, Bruno J . 403.1
Murdoch, Alison 404.1 Vassena, Rita 402.3
Nagaki, Miyuki P.056 Veiga, Anna 102.2, 400.3
Nardo, Luciano 204.3 Velleman, Ramon 308.1
Naumova, Anna K. 309.1 Verhaak, Chris 409.2
Nazzaro, A. P.057 Wakimoto, Eiko P.074
Nouri, Mohammad P.093 Watrelot, Antoine 202.1
Nozha, Chakroun Feki 207.1 Wenger, Jean-Marie 100.5
Okubo, Tsuyoshi 410.5 Wolman, Igal 312.5
Ortega, Carolina P.060, P.061 Woodruff, Teresa K. 200.3
Osada, Hisao 407.4 Yang, Seong-Ho P.075
Pai, Hrishikesh 410.4 Yaron, Yuval 311.1
Palermo, Gianpiero 306.4, 406.4 Yelian, Frank 304.5
Peluffo, Marina C. P.062 Yildirim, Atilla P.076
Peters, Kathleen P.063 Yong, Zeng P.053
Petignat, Patrick 204.1 Yoshida, Atsumi 307.4
Pouly, J. Luc 208.2 Yoshida, Hiroaki 304.2
Prasad, Sudha 301.5 Younas, Kinza 211.3, 211.7
Qiao, Jie 202.4 Young, Lorraine 203.1
Raine-Fenning, Nicholas 103.1, 202.3 Zalel, Yuron 103.3, 202.2
Rajaei, Farzad P.095 Zech, Herbert 308.4
Rajesh, Hemashree P.064 Zhang, John 303.3
Remohí Gimenez, José 408.1 Zorn, Branko 406.3
Roux, Christophe 312.2
Ruvalcaba, Luis Arturo P.065
Salerno, Annalisa P.066
Sallam, Hassan 401.2
Salman, Tarique 410.7, P.067
Santi, Alessandro 309.7
Schenker, Joseph 301.1
Sermon, Karen 101.4
Shoham, Zeev 201.1
Shozu, Makio 304.4
Silber, Sherman 305.2
Smirnova, Anna 410.2
Smith, Gary 306.1
Smith, Venessa 207.3
Son, Weon-Young 310.3
Stadtmauer, Laurel 309.5
Steiner, Hans-Peter 207.4
Streuli, I. 206.4
Swain, Jason 207.5
Taha, Tamer Fouad 312.1
Takefman, Janet 409.4
Tan, Seang Lin 104.1, 300.3, 305.4
Tanaka, Atsushi 308.2
Tang-Pedersen, Mikael 312.6, P.069
Tavmergen, Erol 210.5
April 19-22, 2009 108

CE R T I F I CAT E OF AT T E NDA NCE
THIS IS TO CERTIFY THAT
HAS ATTENDED THE
15TH WORLD CONGRESS ON IN VITRO FERTILIZATION | 4TH WORLD CONGRESS ON IN VITRO MATURATION
GENEVA, SWITZERLAND – APRIL 19-22, 2009
Takahide Mori Jean-Bernard Dubuisson Seang Lin Tan

President Congress President President
International Society for In Vitro Fertilization International Society for In Vitro Maturation

Isivf09 Brochure Final

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SPONSOR ACKNOWLEDGEMENT

i 8 Map / Floor Plans

TIMUR GURGAN S. VAN DER POEL

C. DE GEYTER PHILIPPE BOUCHARD MILTON LEONG

April 19-22, 2009 2

WELCOME FROM THE PRESIDENT OF ISIVF

Dear Colleagues, Ladies and Gentlemen,

I wish you a wonderful Congress and a memorable stay in Geneva!

1 Promulgating ethical practice and standards of practice of IVF treatment;

WELCOME FROM THE PRESIDENT OF ISIVM

On behalf of the Organizing and Scientific Committees it

However, the learning process is ongoing and keeping

April 19-22, 2009 4

WELCOME FROM THE CONGRESS PRESIDENT

Geneva is an international business and art centre. It is

April 19-22, 2009 6

• Delegate Bag Luggage

CCV Gare de Cornavin

April 19-22, 2009 8

Location of Room 18 (Level -1)

About Geneva Currency

April 19-22, 2009 10

Electricity Taxes and Tax Refund

Languages 3 easy steps to claiming your refund in Switzerland

Shopping THROUGH CUSTOMS:

SOCIAL PROGRAMME &

Dress Code: Casual

Accompanying Persons Tour

April 19-22, 2009 12

April 19-22, 2009 14

INSTRUCTIONS FOR ORAL

SUNDAY, APRIL 19 14:00-20:00

Mounting time: MONDAY, APRIL 20 07:00-08:30

April 19-22, 2009 16

April 19-22, 2009 18

Merck Serono Satellite Symposium GONADOTROPINS FUNCTIONAL PROJECT I:

CHAIR: Marc Germond (Switzerland)

MOLECULAR ACTION OF LH ON THE FOLLICLE

THE ROLE OF rFSH/rLH IN FOLLICULAR DEVELOPMENT

CLINICAL EXPERIENCE WITH A NEW COMBINATION OF

IBSA Satellite Symposium HELPING YOU TO IMPROVE YOUR SUCCESS

DOES THE hCG DOSE FOR FINAL FOLLICULAR

ARE THESE DIFFERENT HMGS?

IVF IN THE REAL WORLD: SOCIAL AND ETHICAL

April 19-22, 2009 20

Course 1: Endometriosis and Infertility Room: Madrid

Course 2: PGD and ART Outcomes Room: Rome

Course 3: Stem Cells and Reprogramming Room: Brussels

12:00-13:00 Lunch Break

13:00-16:30 Afternoon Courses

Course 4: Ultrasonography and ART Room: Madrid

Course 5: Germ Cell and Tissue Ovarian

Course 6: Low Cost IVF Room: Brussels

08:30-10:15 Plenary I Room: Salle 2

10:15-10:45 Coffee Break, Exhibits and Poster Viewing

10:45-12:30 Concurrent Symposium 1 - Implantation Room: Salle 3

Co-Chairs: Bischof, Paul - Geneva, Switzerland

10:45-12:30 Concurrent Symposium 2 - Ultrasound and Controlled Room: Salle 2

Co-Chairs: Raine-Fenning, Nicholas - Nottingham, UK

April 19-22, 2009 22

10:45-12:30 ALPHA Session (Scientists in Reproductive Medicine) Room: Salle 4

Co-Chairs: Handyside, Alan - London, UK

203.1 EPIGENETICS AND SAFETY IN ART