UNIVERSITY OF SANTO TOMAS

FACULTY OF PHARMACY | DEPARTMENT OF MEDICAL TECHNOLOGY

IMMUNOHEMATOLOGY and TRANSFUSION MEDICINE
Blood Components and Transfusion

Components – parts of whole blood that can be separated by centrifugation

 Red blood cells

 Plasma, and its derivatives
 Cryoprecipitated antihemophilic factor (AHF)
 Platelets

Blood Collection

 Collected in a primary bag with anticoagulant-preservative mixture

o Closed system – entire blood collection set is sterile, consisting of primary bag
with attached satellite bags and tubings
o Open system – administration ports or other areas are exposed to air, and
allowable storage time is reduced due to potential bacterial contamination

 Anticoagulant-preservative mixture – 63 mL or 70 mL depending on blood bag

o ACD-A (Acid Citrate-dextrose formula A) – 21 days,
 Can be used for APHERESIS
o CPD (Citrate-phosphate dextrose) – 21 days
o CP2D (Citrate-phosphate-double-dextrose) – 21 days
o CDPA-1 (Citrate-phosphate-dextrose-adenine) – 35 days

 Additive solutions – enhances red cell survival and function after plasma separation
o AS-1 (Adsol) and AS-5 (Nutricel) – 42 days
 Dextrose, adenine, mannitol, saline
o AS-3 (Optisol) – 42 days
 Dextrose, adenine, saline, citrate

o 100 mL for 450 mL blood unit

o 110 mL for 500 mL blood unit
o Should be added within 72 hours after whole blood collection

 Dextrose – Supports ATP generation by glycolytic pathway

 Adenine – Substrate for red cell ATP synthesis
 Citrate – Chelates calcium, protects red cell membrane
 Sodium biphosphate – prevents excessive decrease in pH
 Mannitol – Membrane stabilizer (osmotic diuretic)

 Rejuvenation solution – extend life span of RBC units for up to 3 days after expiry
o Contains pyruvate, inosine, phosphate, and adenine
o Wash the unit before transfusion in order to remove inosine (toxic)

 Whole blood collection volume

o For 63 mL mixture: 450 mL  45 mL of blood
o For 70 mL mixture: 500 mL  50 mL of blood
o Reduce mixture for blood collection of less than 300 mL

 “Red Blood Cells Low Volume”

o 300 to 404 mL of whole blood collected for 450 mL blood unit
o 333 to 449 mL of whole blood collected for 500 mL blood unit
o Platelets, FFP, and cAHF should not be prepared

Blood Component Preparation and Storage

 Storage and time is crucial

o Room temperature (not below 20 oC) first when platelets will be prepared
o Refrigerate whole blood at 1-6 oC when platelets will not be prepared

 Separate components by CENTRIFUGATION

o Variables may include:
 Speed of centrifuge at RPM
 Length of time of centrifuge
 Light spin – Short centrifugation time at low RPM
 Heavy spin – Long centrifugation time at higher RPM

Bloodbank | 1
UNIVERSITY OF SANTO TOMAS
FACULTY OF PHARMACY | DEPARTMENT OF MEDICAL TECHNOLOGY
Whole Blood

 Hematocrit: 38%
 Platelets, white cells and labile clotting factors do not survive
 Used to make components of blood

 Storage Temperature: 1-6 oC

 Expiration
o CPD, CP2D – 21 days; CPDA-1 – 35 days

Red Blood Cells

 Prepared by centrifugation or sedimentation, or obtained by apheresis

 Preferred to be prepared after donation
o To manufacture platelet concentrates, frozen plasma, or cryoprecipitate
within 8 hours of collection
 Storage Temperature: 1-6 oC
 Expiration:
o CPD, CP2D – 21 days; CPDA-1 – 35 days
 CPDA-1: 200-250 mL plasma can be removed
 RBC: 65-80% Hematocrit
o With additive solution: 42 days
 Additional 50 mL plasma can be removed
 Due to 150 mL adenine-saline added back to cells
 RBC: 55-65% Hematocrit
 Final Red Cell Volume: 160-275 mL
 Final Hemoglobin: 50-80 grams
 QC: Hematocrit:  80% for CPDA-1 units

Red Blood Cell Aliquots

o Transfused during neonatal period or infants younger than 4 months age

o Indications:
 Anemia caused by spontaneous fetomaternal or fetoplacental
hemorrhage
 Twin-twin transfusion
 Obstetric accidents
 Internal hemorrhage
 Iatrogenic anemia – 10& blood volume removed for lab tests

o 10-25 mL of RBC is usually need

 Precise volume of blood desired can be aspirated into a syringe
through a large-bore needed inserted through and injection site
coupler
 Should be closed securely with a sterile cap
 Label with the following:
o Expiration date
o Patient identification
o Volume transfused
o Preservative
o ABO type

o Initial testing for neonates:

 ABO, Rh, antibody screening
 Use serum or plasma from the infant or mother
 AABB Standards:
 Repeat ABO and Rh may be omitted for the remainder
of the neonate’s hospital admission
 Initial RBC antibody screen is negative  Crossmatching
is not required for initial or subsequent transfusion
 Initial antibody screen is positive for clinically significant
RBC antibodies  Neonate must receive blood without
the corresponding antigen or compatible with AHG
phase

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UNIVERSITY OF SANTO TOMAS
FACULTY OF PHARMACY | DEPARTMENT OF MEDICAL TECHNOLOGY
o Expiration: 24 hours
o Storage Temperature: 1-6 oC
o Anticoagulant: CPDA-1
o Transfuse 10 mL/kg with Hct of 80%  Increase Hgb of 3 g/dL
o Contraindication: Blood units with additive solutions

Frozen Red Blood Cells

 Since 1950s: Freezing using glycerol

 Can be used for:
o Patients with rare phenotypes
o Autologous use
o Military to maintain blood inventories

 Cryoprotective agents are used:

o Penetrating Agent
 Involves small molecules that cross the cell membrane into the
cytoplasm  prevents water from migrating outward as
extracellular ice is formed  prevents intracellular dehydration
 Example: Glycerol

o Nonpenetrating Agent
 Involves large molecules that do not enter the cell but from a
shell around it  prevents loss of water and subsequent
dehydration
 Used to freeze hematopoietic progenitor cells
 Example: Hydroxyethyl starch (HES), dimethylsulfoxide (DMSO)

 Two procedures used for freezing RBCs:

o High-glycerol (40% Weight per Volume)
 Increases the cryoprotective power of the glycerol
 Uses a mechanical freezer: -80 oC
 Requires large volume of wash solution for deglycerolization
 Freeze RBCs within 6 days of collection (CPD or CPDA-1) or
within 42 days of collection (AS-1, AS-3, AS-5)
 AABB: Place RBCs in freezer within 4 hours of opening the
system, and freeze donor serum sample for additional tests

o Low-glycerol (20% Weight per Volume)

 Liquid nitrogen is routinely used
 Storage Temperature:  -120 oC
 Temperature fluctuation during storage can cause RBC
destruction

 Storage Temperature:  -65 oC

 Expiration: 10 years
 QC:
o Monitor refrigerators, freezers, water baths, dry thaw baths, centrifuges
o 80% RBC recovery, 70% RBC viability at 24 hours post-transfusion, and
1% residual intracellular glycerol is removed

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UNIVERSITY OF SANTO TOMAS
FACULTY OF PHARMACY | DEPARTMENT OF MEDICAL TECHNOLOGY
Deglycerolized or Washed Red
Blood Cells

 Free of leukocytes, platelets,

and plasma due to washing
system
 Can be used for:
o Patients with PNH
o Patients with IgA
deficiency with
circulation anti-IgA

 Two procedures used for

thawing RBCs:
o High-glycerol (40%
Weight per Volume)
 Immerse
units into a
37 oC water
bath and
wash RBCs
with solutions
of decreasing osmolarity
 12% NaCl  1.6% NaCl  0.9% NaCl + 0.2% dextrose
o Omit 1.6% NaCl for donor blood units with sickle
cell trait  hemolysis may happen in hypertonic
solutions
 Considered as open system

 Storage Temperature: 1-6 oC

 Expiration: 24 hours due to open system
 QC: Do visual hemoglobin check, and Box 13-7

Irradiated Red Blood Cells

 Recipients include:
o Immunocompromised patients
o Patients receiving bone marrow or stem cell transplant
o Fetus undergoing intrauterine transfusion
o Recipients of blood from relatives

 Irradiation
o Inhibits proliferation of T cells and subsequent transfusion-associated
graft-versus-host disease
o Performed using cesium-137 or cobalt-60
o Place radiochromic film label to the component before placing into the
metal canister of the irradiator
 Success: Darkening of film

 Minimum dose of gamma irradiation (FDA and AABB)

o 25 Gy: Central portion of blood unit
o No less than 15 Gy: Any part of the blood unit

 Storage Temperature: 1-6 oC

 Expiration: 28 days from irradiation or original expiry, whichever is first
 QC: Irradiator QC applied 2500 cGy in center of unit

Leukocyte Reduced Red Blood Cells

 AABB: Absolute WBC count: <5 x 106 and at least 85% original RBC mass

 Two major categories of leuko-reduced RBCs:

o Prestorage leukoreduction
 Usage of special filters: Multiple layers of polyester or cellulose
acetate nonwoven fibers are placed in order to trap leukocytes
and platelet  99.9% removal of leukocytes
 Impetus: Biological response modifiers are released from
leukocytes that can promote febrile transfusion reactions

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UNIVERSITY OF SANTO TOMAS
FACULTY OF PHARMACY | DEPARTMENT OF MEDICAL TECHNOLOGY
 Proinflammatory cytokines
 Complement fragments
 Three methods:
 In-line filter – attached to whole blood units, filtered by
gravity  RBCs and plasma can be prepared
 Plasma is initial removed, then packed cells are pass
through an in-line reduction filter
 Sterile docking device is used to attach a leukocyte
reduction ilter to a unit of RBCS  filtered by gravity
 First two methods cannot be used to prepare random-
donor platelets since they are trapped in filters

o Poststorage leukoreduction
 Leukocytes are removed in blood bank prior to issuance of
blood, or at the bedside before transfusion
 Centrifugation: Absolute WBC count is <5 x 108
 Third generation filters: Absolute WBC count is <5 x 106
 Can prevent reactions caused by leukocyte antibodies in
patient’s plasma and leukocytes present transfused blood, but
cannot prevent reactions caused by BRMs

 Frozen, thawed, deglycerolized, and washed RBCs also produced leukoreduced

products (Arnaud and Meryman)
o Remove buffy coat at time of collection after freezing and
deglycerolization could be a possible alternative to leukocyte filtration
after freezing
o Shelf-life: 24 hours

 Useful in trying to avoid the following:

o Febrile nonhemolytic transfusion reactions
o Transfusion-related acute lung injury
o Transmission of Epstein-Barr virus, cytomegalovirus, and human T-cell
lymphotrophic virus

 Storage Temperature: 1-6 oC

 Expiration:
o CPD, CP2D – 21 days; CPDA-1 – 35 days
o With additive solution: 42 days
 QC: Hematocrit:  80% for CPDA-1 units

Plasma Frozen within 24 hours (PF24)

 Storage Temperature:  -18 oC

 Expiration: 1 year

Fresh Frozen Plasma (FFP)

 Storage Temperature:  -18 oC or  -65 oC

 Expiration: 1 year or 7 years, respectively

FFP or PF24 that is thawed

 Storage Temperature: 1-6 oC

 Expiration: 24 hours

Thawed Plasma

 Storage Temperature: 1-6 oC

 Expiration: 5 days from thawing
 QC: NOT LICENSED

Cryoprecipitated AHF

 Storage Temperature:  -18 oC

 Expiration: 1 year
 QC: Factor VIII:  80 IU and 150 mg fibrinogen

Pooled CRYO

 Storage Temperature: 20-24 oC

 Expiration: 4 hours

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UNIVERSITY OF SANTO TOMAS
FACULTY OF PHARMACY | DEPARTMENT OF MEDICAL TECHNOLOGY
 QC: Factor VIII:  80 IU and 150 mg fibrinogen times number in pool

Platelet Concentrates

 Produced during the routine conversion of whole blood or by apheresis

o Apheresis platelets: High yield and minimal RBC contamination
o Whole blood used must be drawn by a single nontraumatic venipuncture
o Prepared within 4 hours of collection

o Whole blood platelets  Random-donor platelets

  5.5 x 1010 platelets
 Plasma: 40 to 70 mL
o Apheresis platelets  Single-donor platelets
  3.0 x 1011 platelets (equivalent to 4-6 RDPs)
 Plasma: Approximately 300 mL

 Can be used for:

o Actively bleeding patients who are thrombocytopenic (<50,000 /uL) due
to decreased production or decreased function
o Patients during radiation and chemotherapy due to induced
thrombocytopenia (<20,000 /uL)
o Thrombocytopenic preoperative patients (<50,000 /uL)
o NOT INDICATED in DIC or ITP
 This is due to increased destruction of platelets in ITP and
increased consumption in DIC

o Single-donor platelets
 Indicated for patients who are unresponsive to RDPs due to HLA
alloimmunization or to limit platelet exposure from multiple
donors

 Storage Temperature:
o RDP: 20-24 oC with continuous agitation
o SDP: 22-24 oC with continuous agitation
 Expiration: 5 days
o Short length due to increased bacterial contamination
o Open system: Within 6 hours
 Quality Control:
o  5.5 x 1010 platelets for RDP
o  3.0 x 1011 platelets for SDP
o pH 6.2

Platelet Aliquots

o For neonates whose counts falls below 50,000 /uL and who are
experiencing bleeding
o Factors that may be associated with thrombocytopenia:
o Immaturity of the coagulation system
o Platelet dysfunction
o Increased platelet destruction
o Dilution effect secondary to massive transfusion or exchange
transfusion
o Intraventricular hemorrhage

o RDP or SDP may be transfused

o Transfuse 5-10 mL/kg  Increase platelet count by 50,000-100,000
o Expiration: 4 hours

Pooled Platelets

 All of the units for a single platelet dose can be pooled into a single bag before
transfusion
o Typically 6-8 units for an adult
o Place a unique pool number on the label

 Storage Temperature: 20-24 oC

 Expiration: 4 hours

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UNIVERSITY OF SANTO TOMAS
FACULTY OF PHARMACY | DEPARTMENT OF MEDICAL TECHNOLOGY
Leukocyte Reduced Apheresis Platelets

 For prevention of febrile nonhemolytic reactions

 Leukoreduction of RDPs using a leukoreduction filter designed for platelets

 Storage Temperature: 20-24 oC

 Expiration: 5 days
 Quality Control:
o <8.3 x 105 leukocytes in 95% of unit for RDPs
o <5 x 106 leukocytes for final pooled platelets
o <5 x 106 leukocytes in 95% of unit for SDPs

Apheresis Granulocytes

 Storage Temperature: 20-24 oC

 Expiration: 24 hours
 QC:  1 x 1010 granulocytes in 75% of units tested

Blood Donor Screening

The Blood Groups

Pretransfusion Testing