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Description

ProductSheet

M1(ATCCCRL2038)

Organism:Musmusculus,transgenicforSV40earlyregion,mouse,transgenicforSV40earlyregion
Tissue:kidney,cortex,collectingduct
CellType:Epithelial
Morphology:epithelial
GrowthProperties:adherent

BatchSpecificInformation
PleasereadthisFIRST
RefertotheCertificateofAnalysisforbatchspecifictestresults.

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
2

SAFETYPRECAUTION
ATCChighlyrecommendsthatprotectiveglovesandclothingalwaysbeusedandafullfacemaskalwaysbe
wornwhenhandlingfrozenvials.Itisimportanttonotethatsomevialsleakwhensubmersedinliquidnitrogen
andwillslowlyfillwithliquidnitrogen.Uponthawing,theconversionoftheliquidnitrogenbacktoitsgas
phasemayresultinthevesselexplodingorblowingoffitscapwithdangerousforcecreatingflyingdebris.

Unpacking&StorageInstructions

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
A1:1mixtureofDulbecco'smodifiedEagle'smedium
andHam'sF12mediumwith2.5mMLglutamine
adjustedtocontain15mMHEPES,0.5mMsodium
pyruvateand1.2g/Lsodiumbicarbonate
supplementedwith0.005mMdexamethasoneand5%
fetalbovineserum

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:M1(ATCCCRL2038)

1. Checkallcontainersforleakageorbreakage.
2. Removethefrozencellsfromthedryicepackagingandimmediatelyplacethecellsatatemperature
below130C,preferablyinliquidnitrogenvapor,untilreadyforuse.

HandlingProcedureforFrozenCells
Toinsurethehighestlevelofviability,thawthevialandinitiatethecultureassoonaspossibleuponreceipt.If
uponarrival,continuedstorageofthefrozencultureisnecessary,itshouldbestoredinliquidnitrogenvapor
phaseandnotat70C.Storageat70Cwillresultinlossofviability.
1. Thawthevialbygentleagitationina37Cwaterbath.Toreducethepossibilityofcontamination,keep
theOringandcapoutofthewater.Thawingshouldberapid(approximately2minutes).
2. Removethevialfromthewaterbathassoonasthecontentsarethawed,anddecontaminateby
dippinginorsprayingwith70%ethanol.Alloftheoperationsfromthispointonshouldbecarriedout
understrictasepticconditions.
3. Transferthevialcontentstoa75cm2tissuecultureflaskanddilutewiththerecommendedcomplete
culturemedium(seethespecificbatchinformationfortherecommendeddilutionratio).Itisimportant
toavoidexcessivealkalinityofthemediumduringrecoveryofthecells.Itissuggestedthat,priorto
theadditionofthevialcontents,theculturevesselcontainingthegrowthmediumbeplacedintothe
incubatorforatleast15minutestoallowthemediumtoreachitsnormalpH(7.0to7.6).
4. Incubatethecultureat37Cinasuitableincubator.A5%CO2inairatmosphereisrecommendedif
usingthemediumdescribedonthisproductsheet.
Note:Itisnotnecessarytoremovethecryoprotectiveagent.Ifitisdesiredthatthecryoprotectiveagentbe
removedimmediately,orthatamoreconcentratedcellsuspensionbeobtained,centrifugethecellsuspension
atapproximately125xgfor5to10minutes.Discardthesupernatantandresuspendthecellswithfresh
growthmediumatthedilutionratiorecommendedinthespecificbatchinformation.

HandlingProcedureforFlaskCultures

AmericanTypeCultureCollection
POBox1549
Manassas,VA20108USA
www.atcc.org
800.638.6597or703.365.2700
Fax:703.365.2750
Email:Tech@atcc.org

Orcontactyourlocaldistributor

Page1of3

Theflaskwasseededwithcells(seespecificbatchinformation)grownandcompletelyfilledwithmediumat
ATCCtopreventlossofcellsduringshipping.
1. Uponreceiptvisuallyexaminethecultureformacroscopicevidenceofanymicrobialcontamination.
Usinganinvertedmicroscope(preferablyequippedwithphasecontrastoptics),carefullycheckfor
anyevidenceofmicrobialcontamination.Alsochecktodetermineifthemajorityofcellsarestill
attachedtothebottomoftheflaskduringshippingtheculturesaresometimeshandledroughlyand
manyofthecellsoftendetachandbecomesuspendedintheculturemedium(butarestillviable).
2. Ifthecellsarestillattached,asepticallyremoveallbut5to10mLoftheshippingmedium.The
shippingmediumcanbesavedforreuse.Incubatethecellsat37Cina5%CO2inairatmosphere
untiltheyarereadytobesubcultured.
3. Ifthecellsarenotattached,asepticallyremovetheentirecontentsoftheflaskandcentrifugeat
125xgfor5to10minutes.Removeshippingmediumandsave.Resuspendthepelletedcellsin10
mLofthismediumandaddto25cm2flask.Incubateat37Cina5%CO2inairatmosphereuntilcells
are

SubculturingProcedure

ProductSheet

M1(ATCCCRL2038)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
2

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
A1:1mixtureofDulbecco'smodifiedEagle'smedium
andHam'sF12mediumwith2.5mMLglutamine
adjustedtocontain15mMHEPES,0.5mMsodium
pyruvateand1.2g/Lsodiumbicarbonate
supplementedwith0.005mMdexamethasoneand5%
fetalbovineserum

Volumesusedinthisprotocolarefor75cm2flaskproportionallyreduceorincreaseamountofdissociation
mediumforculturevesselsofothersizes.
1. Removeanddiscardculturemedium.
2. Brieflyrinsethecelllayerwith0.25%(w/v)Trypsin0.53mMEDTAsolutiontoremovealltracesof
serumthatcontainstrypsininhibitor.
3. Add2.0to3.0mLofTrypsinEDTAsolutiontoflaskandobservecellsunderaninvertedmicroscope
untilcelllayerisdispersed(usuallywithin5to15minutes).
Note:Toavoidclumpingdonotagitatethecellsbyhittingorshakingtheflaskwhilewaitingforthe
cellstodetach.Cellsthataredifficulttodetachmaybeplacedat37Ctofacilitatedispersal.
4. Add6.0to8.0mLofcompletegrowthmediumandaspiratecellsbygentlypipetting.
5. Addappropriatealiquotsofthecellsuspensiontonewculturevessels.
6. Incubateculturesat37C.
SubcultivationRatio:Asubcultivationratioof1:3to1:4isrecommended
MediumRenewal:2to3timesperweek
Note:FormoreinformationonenzymaticdissociationandsubculturingofcelllinesconsultChapter10in
CultureofAnimalCells,amanualofBasicTechniquebyR.IanFreshney,3rdedition,publishedby
AlanR.Liss,N.Y.,1994.

CryopreservationMedium
Completeculturemediumdescribedabovesupplementedwith5%(v/v)DMSO.CellculturetestedDMSOis
availableasATCCCatalogNo.4X.

Comments
Thecellsretainmanycharacteristicsofcorticalcollectingduct(CCD)cellsincludingmorphologyandCCD
antigens.
MostcelllinesclonedfromM1exhibitcharacteristicsofeitherintercalatedcells(ICC)orprinciplecells(PC)of
theCCD.
5to10%ofthecellsexhibitadualPCICCphenotype.
Whengrownonpermeablesupports,thecellsdevelopalumennegativetransepithelialpotentialdifference.

References
Referencesandotherinformationrelatingtothisproductareavailableonlineatwww.atcc.org.

CitationofStrain

BiosafetyLevel:2
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:M1(ATCCCRL2038)

Appropriatesafetyproceduresshouldalwaysbeusedwiththismaterial.Laboratorysafetyisdiscussedin
thecurrentpublicationoftheBiosafetyinMicrobiologicalandBiomedicalLaboratoriesfromtheU.S.
DepartmentofHealthandHumanServicesCentersforDiseaseControlandPreventionandNationalInstitutes
forHealth.

ATCCWarranty
TheviabilityofATCCproductsiswarrantedfor30daysfromthedateofshipment,andisvalidonlyifthe
productisstoredandculturedaccordingtotheinformationincludedonthisproductinformationsheet.ATCC
liststhemediaformulationthathasbeenfoundtobeeffectiveforthisstrain.Whileother,unspecifiedmedia
mayalsoproducesatisfactoryresults,achangeinmediaortheabsenceofanadditivefromtheATCC
recommendedmediamayaffectrecovery,growthand/orfunctionofthisstrain.Ifanalternativemedium
formulationisused,theATCCwarrantyforviabilityisnolongervalid.

Disclaimers
AmericanTypeCultureCollection
POBox1549
Manassas,VA20108USA
www.atcc.org
800.638.6597or703.365.2700
Fax:703.365.2750
Email:Tech@atcc.org

Orcontactyourlocaldistributor

Page2of3

Thisproductisintendedforlaboratoryresearchpurposesonly.Itisnotintendedforuseinhumans.
WhileATCCusesreasonableeffortstoincludeaccurateanduptodateinformationonthisproductsheet,
ATCCmakesnowarrantiesorrepresentationsastoitsaccuracy.Citationsfromscientificliteratureand
patentsareprovidedforinformationalpurposesonly.ATCCdoesnotwarrantthatsuchinformationhasbeen
confirmedtobeaccurate.
Thisproductissentwiththeconditionthatyouareresponsibleforitssafestorage,handling,anduse.ATCC
isnotliableforanydamagesorinjuriesarisingfromreceiptand/oruseofthisproduct.Whilereasonableeffort
ismadetoinsureauthenticityandreliabilityofstrainsondeposit,ATCCisnotliablefordamagesarisingfrom
themisidentificationormisrepresentationofcultures.

PleaseseetheenclosedMaterialTransferAgreement(MTA)forfurtherdetailsregardingtheuseofthis
product.TheMTAisalsoavailableonourWebsiteatwww.atcc.org
AdditionalinformationonthiscultureisavailableontheATCCwebsiteatwww.atcc.org.
ATCC2013.Allrightsreserved.ATCCisaregisteredtrademarkoftheAmericanTypeCultureCollection.[07/02]

ProductSheet

M1(ATCCCRL2038)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
2

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
A1:1mixtureofDulbecco'smodifiedEagle'smedium
andHam'sF12mediumwith2.5mMLglutamine
adjustedtocontain15mMHEPES,0.5mMsodium
pyruvateand1.2g/Lsodiumbicarbonate
supplementedwith0.005mMdexamethasoneand5%
fetalbovineserum

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:M1(ATCCCRL2038)

AmericanTypeCultureCollection
POBox1549
Manassas,VA20108USA
www.atcc.org
800.638.6597or703.365.2700
Fax:703.365.2750
Email:Tech@atcc.org

Orcontactyourlocaldistributor

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