1. II. Blood and BloodII. Blood and Blood
ComponentsComponents
Terry Kotrla, MS, MT(ASCP)BBTerry Kotrla, MS, MT(ASCP)BB
Spring 2010Spring 2010

2. Goals Of Blood CollectionGoals Of Blood Collection
 Maintain viability and functionMaintain viability and function
 Prevent physical changesPrevent physical changes
 Minimize bacterial contaminationMinimize bacterial contamination

3. Anticoagulants Preservative
Solutions
 Anticoagulants prevent blood clottingAnticoagulants prevent blood clotting
 Preservatives provide nutrients for cellsPreservatives provide nutrients for cells
 HeparinHeparin
– Rarely if ever used anymoreRarely if ever used anymore
– Anticoagulant ONLYAnticoagulant ONLY
– Transfuse within 48 hours, preferably 8Transfuse within 48 hours, preferably 8

4. AnticoagulantsAnticoagulants
CPDCPD CPD-A1CPD-A1
Storage timeStorage time 21 days21 days 35 days35 days
TemperatureTemperature 1-6 C1-6 C 1-6 C1-6 C
Slows glycolytic activitySlows glycolytic activity
AdenineAdenine NoneNone Substrate for ATP synthesisSubstrate for ATP synthesis
VolumeVolume 450 +/- 10%450 +/- 10%
DextroseDextrose Supports ATP generation by glycolyticSupports ATP generation by glycolytic
pathwaypathway
CitrateCitrate Prevents coagulation by binding calciumPrevents coagulation by binding calcium

5. Additive SolutionAdditive Solution
 Primary bag with satellite bags attached.Primary bag with satellite bags attached.
 One bag has additive solution (AS)One bag has additive solution (AS)
 Unit drawn into CPD anticoagulantUnit drawn into CPD anticoagulant

6. Additive SolutionAdditive Solution
 Remove platelet rich plasma within 72 hoursRemove platelet rich plasma within 72 hours
 Add additive solution to RBCs, ADSOL, whichAdd additive solution to RBCs, ADSOL, which
consists of:consists of:
– SalineSaline
– AdenineAdenine
– GlucoseGlucose
– MannitolMannitol
 Extends storage to 42 daysExtends storage to 42 days
 Final hematocrit approximately 66%Final hematocrit approximately 66%

7. Changes Occur During StorageChanges Occur During Storage
 Shelf life = expiration dateShelf life = expiration date
– At end of expiration must have 75% recoveryAt end of expiration must have 75% recovery
– At least 75% of transfused cells remain inAt least 75% of transfused cells remain in
circulation 24 hours AFTER transfusioncirculation 24 hours AFTER transfusion

8. Storage LesionStorage Lesion
 Biochemical changes which occur at 1-6CBiochemical changes which occur at 1-6C
 Affects oxygen dissociation curve, increasedAffects oxygen dissociation curve, increased
affinity of hemoglobin for oxygen.affinity of hemoglobin for oxygen.
– Low 2,3-DPG, increased OLow 2,3-DPG, increased O22 affinity, less Oaffinity, less O22 released.released.
– pH drops causes 2,3-DPG levels to fallpH drops causes 2,3-DPG levels to fall
– Once transfused RBCs regenerate ATP and 2,3-DPGOnce transfused RBCs regenerate ATP and 2,3-DPG
 Few functional platelets presentFew functional platelets present
 Viable (living) RBCs decreaseViable (living) RBCs decrease

9. Plasma
hemoglobin
Plasma K+
Viable cells
pH
ATP
2,3-DPG
Plasma Na+
Helps release oxygen
from hemoglobin (once
transfused, ATP & 2,3-
DPG return to normal)
K+Na+

10. Storage LesionStorage Lesion
 Significant for infants and massiveSignificant for infants and massive
transfusion.transfusion.
 Other biochemical changesOther biochemical changes
– ATP decreasesATP decreases
– Potassium increasesPotassium increases
– Sodium decreasesSodium decreases
– Plasma hemoglobin increasesPlasma hemoglobin increases

11. Preparation of ComponentsPreparation of Components
 Collect unit within 15 minutes to preventCollect unit within 15 minutes to prevent
activation of coagulation systemactivation of coagulation system
 Draw into closed system – primary bag withDraw into closed system – primary bag with
satellite bags with hermetic seal between.satellite bags with hermetic seal between.
 If hermetic seal broken transfuse within 24 hoursIf hermetic seal broken transfuse within 24 hours
if stored at 1-4C, 4 hours if stored at 20-24Cif stored at 1-4C, 4 hours if stored at 20-24C

12. Preparation of ComponentsPreparation of Components
 Centrifuge – light spin, platelets suspendedCentrifuge – light spin, platelets suspended
 Remove platelet rich plasma (PRP)Remove platelet rich plasma (PRP)
 Centrifuge PRP heavy spinCentrifuge PRP heavy spin
 Remove platelet poor plasmaRemove platelet poor plasma
 Freeze plasma solid within 8 hoursFreeze plasma solid within 8 hours
 Thaw plasma at 1-4C – precipitate formsThaw plasma at 1-4C – precipitate forms
 Centrifuge, express plasma leavingCentrifuge, express plasma leaving
cryoprecipitate. Store both at -18Ccryoprecipitate. Store both at -18C
 RBCs – CPD – 21 days, ADSOL – 42 days – 1-RBCs – CPD – 21 days, ADSOL – 42 days – 1-
6C6C

14. Preparation of ComponentsPreparation of Components
 Summary – One unit of whole blood canSummary – One unit of whole blood can
produce:produce:
– Packed RBCsPacked RBCs
– Fresh frozen plasma (FFP)Fresh frozen plasma (FFP)
– Cryoprecipitate (CRYO)Cryoprecipitate (CRYO)
– Single donor plasma (SDP) – cyro removedSingle donor plasma (SDP) – cyro removed
– Platelets – terms PC (platelet concentrate) ORPlatelets – terms PC (platelet concentrate) OR
RD PC (random donor platelet concentrate)RD PC (random donor platelet concentrate)

15. Preparation of ComponentsPreparation of Components
 Sterile docking device joins tubingSterile docking device joins tubing
– Used to add satellite bags to maintain originalUsed to add satellite bags to maintain original
expiration of componentexpiration of component
– May be used to pool componentsMay be used to pool components

16. Blood Component General InformationBlood Component General Information
 Blood separated into components toBlood separated into components to
specifically treat patients with productspecifically treat patients with product
neededneeded
 Advantages of component separationAdvantages of component separation
– Allow optimum survival of each componentAllow optimum survival of each component
– Transfuse only component neededTransfuse only component needed

17. Blood Component General InformationBlood Component General Information
 Transfusion practiceTransfusion practice
– Transfusion requires doctor’s prescriptionTransfusion requires doctor’s prescription
– All components MUST be administeredAll components MUST be administered
through a filterthrough a filter
– Infuse quickly, within 4 hoursInfuse quickly, within 4 hours
– D (Rh) neg require D neg cellular productsD (Rh) neg require D neg cellular products
– ABO identical preferred, ABO compatible OKABO identical preferred, ABO compatible OK
– ““Universal donor” – RBCs group O, plasmaUniversal donor” – RBCs group O, plasma
ABAB

18. Blood Component General InformationBlood Component General Information
 Fresh Whole BloodFresh Whole Blood
– Blood not usually available until 12-24 hoursBlood not usually available until 12-24 hours
– CandidatesCandidates
 Newborns needing exchange transfusionNewborns needing exchange transfusion
 Patients requiring leukoreduced productsPatients requiring leukoreduced products

19. Blood Component General InformationBlood Component General Information
 Summary of storage temperatures:Summary of storage temperatures:
– Liquid RBCs 1-6CLiquid RBCs 1-6C
– Platelets, Cryo (thawed) and granulocytes 20-Platelets, Cryo (thawed) and granulocytes 20-
24C (room temperature)24C (room temperature)
– ANY frozen plasma product ≤ -18CANY frozen plasma product ≤ -18C
– ANY liquid plasma product EXCEPT Cryo 1-ANY liquid plasma product EXCEPT Cryo 1-
6C6C

20. Blood ComponentsBlood Components
 CellularCellular
– Red blood cell productsRed blood cell products
– PlateletsPlatelets
– GranulocytesGranulocytes
 PlasmaPlasma
– FFPFFP
– CryoprecipitateCryoprecipitate

21. Products With Red CellsProducts With Red Cells

22. Whole BloodWhole Blood
 Clinical indications for use of WB are extremely limited.
 Used for massive transfusion to correct acute
hypovolemia such as in trauma and shock, exchange
transfusion.
 RARELY used today, platelets non-functional, labile
coagulation factors gone.
 Must be ABO identical.

23. Changes in Stored BloodChanges in Stored Blood

24. Red Blood Cells, PackedRed Blood Cells, Packed
(PRBC)(PRBC)
 Used to treat symptomatic anemia and routine
blood loss during surgery
 Hematocrit is approximately 80% for non-
additive (CPD), 60% for additive (ADSOL).
 Allow WB to sediment or centrifuge WB, remove
supernatant plasma.

25. Leukocyte Reduced Red Cells (LR-Leukocyte Reduced Red Cells (LR-
RBC)RBC)
 Leukocytes can induce adverse affects during transfusion, primarily
febrile, non-hemolytic reactions.
 Reactions to cytokines produced by leukocytes in transfused units.
 Other explanations to reactions include: immunization of recipient to
transfused HLA or granulocyte antigens, micro aggregates and
fragmentation of granulocytes.
 Historically, indicated only for patients who had 2 or more febrile
transfusion reactions, now a commonly ordered, popular
component.
 “CMV” safe blood, since CMV lives in WBCs.
 Most blood centers now leukoreduce blood immediately after
collection.
 Bed side filters are available to leukoreduce products during
transfusion.

26. Leukocyte ReductionLeukocyte Reduction

27. Washed Red Blood Cells (W-
RBCs)
 Washing removes plasma proteins, platelets, WBCs and
micro aggregates which may cause febrile or urticarial
reactions.
 Patient requiring this product is the IgA deficient patient
with anti-IgA antibodies.
 Prepared by using a machine which washes the cells 3
times with saline to remove and WBCs.
 Two types of labels:
– Washed RBCs - do not need to QC for WBCs.
– Leukocyte Poor WRBCs, QC must be done to guarantee
removal of 85% of WBCs. No longer considered effective
method for leukoreduction.
 e. Expires 24 hours after unit is entered.

28. Cell Washer to Prepare WashedCell Washer to Prepare Washed
CellsCells

29. Frozen BloodFrozen Blood

30. Red Blood Cells Frozen; Red Blood Cells
Deglycerolized (D-RBC)
 Blood is frozen to preserve: rare types, for autologous
transfusion, stock piling blood for military mobilization
and/or civilian natural disasters.
 Blood is drawn into an anticoagulant preservative.
– Plasma is removed and glycerol is added.
– After equilibration unit is centrifuged to remove excess glycerol
and frozen.
 Expiration
– If frozen, 10 years.
– After deglycerolization, 24 hours.
 Storage temperature
– high glycerol -65 C.
– low glycerol -120 C, liquid nitrogen.

31. Red Blood Cells Frozen; Red Blood Cells
Deglycerolized (D-RBC)
 Thaw unit at 37C, thawed RBCs will have high
concentration of glycerol.
 A solution of glycerol of lesser concentration of the
original glycerol is added.
 This causes glycerol to come out of the red blood cells
slowly to prevent hemolysis of the RBCs.
 After a period of equilibration the unit is spun, the
solution is removed and a solution with a lower glycerol
concentration is added.
 This procedure is repeated until all glycerol is removed,
more steps are required for the high glycerol stored
units.
 The unit is then washed.

32. Rejuvenated Red Blood Cells
 A special solution is added to expired RBCs up
to 3 days after expiration to restore 2,3-DPG and
ATP levels to prestorage values.
 Rejuvenated RBCs regain normal characteristics
of oxygen transport and delivery and improved
post transfusion survival.
 Expiration is 24 hours or, if frozen, 10 years

33. Platelet ProductsPlatelet Products

34. Platelets (PLTS), Platelet Concentrate (PC) or
Random Donor Platelet Concentrate (RD-PC)
 Used to prevent spontaneous bleeding or stop
established bleeding in thrombocytopenic patients.
 Prepared from a single unit of whole blood.
 Due to storage at RT it is the most likely component to
be contaminated with bacteria.
 Therapeutic dose for adults is 6 to 10 units.
 Some patients become "refractory" to platelet therapy.
 Expiration is 5 days as a single unit, 4 hours if pooled.
 Store at 20-24 C (RT) with constant agitation.
 D negative patients should be transfused with D negative
platelets due to the presence of a small number of
RBCs.

35. Preparation of plateletPreparation of platelet
concentrateconcentrate
RBCs PRP
Plasma
Platelet
concentrate

36. Platelets (PLTS), Platelet Concentrate (PC) or
Random Donor Platelet Concentrate (RD-PC)
 One bag from ONE donorOne bag from ONE donor
 Need 6-10 for therapeutic doseNeed 6-10 for therapeutic dose

37. Pooling PlateletsPooling Platelets
 6-10 units transferred into one bag6-10 units transferred into one bag
 Expiration = 4 hoursExpiration = 4 hours

38. Platelets Pheresis, Apheresis Platelet Concentrate, Single
Donor Platelet Concentrate (SD-PC)
 Used to decrease donor exposure, obtain HLA matched
platelets for patients who are refractory to RD-PC or
prevent platelet refractoriness from occurring.
 Prepared by hemapheresis, stored in two connected
bags to maintain viability.
 One pheresed unit is equivalent to 6-8 RD-PC.
 Store at 20-24 C (RT) with agitation for 5 days, after
combining, 24 hours
 D negative patients should be transfused with D negative
platelets due to the presence of a small number of RBCs

39. ApheresisApheresis

40. ApheresisApheresis

41. Platelets Pheresis
 One bag (unit) fromOne bag (unit) from
one donorone donor
 One unit is aOne unit is a
therapeutic dosetherapeutic dose
 VolumeVolume
approximately 250approximately 250
ccsccs

42. GranulocytesGranulocytes
Lymphocyte Monocyte
Neutrophils Eosinophils Basophils

43. Granulocytes
 Primary use is for patients with neutropenia who have
gram negative infections documented by culture, but are
unresponsive to antibiotics.
 Therapeutic efficacy and indications for granulocyte
transfusions are not well defined.
 Better antimicrobial agents and use of granulocyte and
macrophage colony stimulating factors best for adults,
best success with this component has been with babies
 Daily transfusions are necessary.
 Prepared by hemapheresis.
 Expiration time is 24 hours but best to infuse ASAP.
 Store at 20-24 C.

44. Plasma ComponentsPlasma Components

45. Fresh Frozen Plasma –Fresh Frozen Plasma –
Volume 200-250ccVolume 200-250cc

46. Fresh Frozen Plasma
(FFP)
 Used to replace labile and non-labile coagulation
factors in massively bleeding patients OR treat
bleeding associated with clotting factor
deficiencies when factor concentrate is not
available.
 Must be frozen within 8 hours of collection.
 Expiration
– frozen - 1 year stored at <-18 C.
– frozen - 7 years stored at <-65 C.thawed - 24 hours

47. Fresh Frozen Plasma
(FFP)
 Storage temperature
– frozen -18 C, preferably -30 C or lower
– thawed - 1-6 C
 Thawed in 30-37C water bath or FDA
approved microwave
 Must have mechanism to detect units
which have thawed and refrozen due to
improper storage.
 Must be ABO compatible

48. Plasma, Liquid Plasma, Recovered
Plasma and Source Plasma
 Used to treat patients with stable clotting factor
deficiencies for which no concentrate is available or for
patients undergoing therapeutic plasmapheresis.
 Prepared by separating the plasma from the RBCs on
or before the 5th day after expiration of the whole
blood.
 Once separated can:
– Freeze, store at -18 C for 5 years
– If not frozen, called liquid plasma, store at 1-6 C for up to 5 days
after expiration of WB.
 Once FFP is one year old can redesignate as Plasma,
expiration is 5 years.

49. Pooled Plasma/Solvent
Detergent Treated
 Most recently licensed product.
 Prepared from pools of no more than 2500 units of ABO
specific plasma frozen to preserve labile coagulation
factors.
 Treated with chemicals to inactivate lipid-enveloped
viruses.
 Contains labile and non-labile coagulation factors but
lacks largest Von Willebrand’s factor multimers.
 Used same as FFP.Safety concerns
– Decreases disease transmission for diseases tested for.
– Doesn’t inactivate viruses with non-lipid envelopes: parvo virus
B19, hepatitis A, and unrecognized pathogens

50. Cryoprecipitate (CRYO), Factor VIII
or Anti-Hemophilic Factor (AHF)
 Cold insoluble portion of plasma that precipitates when
FFP is thawed at 1-6C.
 Cryoprecipitate contains high levels of Factor VIII and
Fibrinogen, used for treatment of hemophiliacs and
Von Willebrands when concentrates are not available.
 Used most commonly for patients with DIC or low
fibrinogen levels.
 A therapeutic dose for an adult is 6 to 10 units.
 Can be prepared from WB which is then designated as
"Whole Blood Cryoprecipitate Removed" or from FFP
– Plasma is frozen.
– Plasma is then thawed at 1-6 C, a precipitate forms.
– Plasma is centrifuged, cryoprecipitate will go to
bottom.
– Remove plasma, freeze within 1 hour of preparation

51. FFP
Frozen
within 8
hours
Thawed
FFP
Cryoprecipitate
(VIII, vW)
Plasma cryoprecipitate, reduced
(TTP, FII, V, Vii, IX, X, XI)
Thaw at 30-37°C
Store at RT 4 hrs
Refrozen with 24 hrs of
separation
Store at ≤18°C 1 yr
5 day expiration at 1-6°C

52. Cryoprecipitate (CRYO), Factor VIII
or Anti-Hemophilic Factor (AHF)
 Storage Temperature
– Frozen -18 C or lower
– Thawed - room temperature
 Expiration:
– Frozen 1 year
– Thawed 6 hours
– Pooled 4 hours
 Best to be ABO compatible but not
important due to small volume

53. Cryoprecipitate – volume 15ccsCryoprecipitate – volume 15ccs

54. Irradiation of Blood Components

55. Irradiation of Blood Components
 Cellular blood components are irradiated to
destroy viable T- lymphocytes which may cause
Graft Versus Host Disease (GVHD).
 GVHD is a disease that results when
immunocompetent, viable lymphocytes in donor
blood engraft in an immunocompromised host,
recognize the patient tissues as foreign and
produce antibodies against patient tissues,
primarily skin, liver and GI tract. The resulting
disease has serious consequences including
death.
 GVHD may be chronic or acute

56. Irradiation of Blood Components
 Patients at greatest risk are:
– severely immunosuppressed,
– immunocompromised,
– receive blood donated by relatives, or
– fetuses receiving intrauterine transfusions
 Irradiation inactivates lymphocytes, leaving platelets,
RBCs and granulocytes relatively undamaged.
 Must be labeled "irradiated".
 Expiration date of Red Blood Cell donor unit changes to
28 days.
 May be transfused to "normal" patients if not used by
intended recipient.

57. Irradiation of Blood Components

58. Donor Blood Inspection and Disposition
 It is required that donor units be inspected periodically
during storage and prior to issuing to patient.
 The following may indicate an unacceptable unit:
– Red cell mass looks purple or clots are visible.
– Zone of hemolysis observed just above RBC mass, look for
hemolysis in sprigs, especially those closest to the unit.
– Plasma or supernatant plasma appears murky, purple, brown or
red.
– A greenish hue need not cause a unit to be rejected.
– Inspect platelets for aggregates.
 Inspect FFP and CRYO for signs of thawing, evidence of
cracks in bag, or unusual turbidity in CRYO or FFP (i.e.,
extreme lipemia).

59. Inspection of Donor BloodInspection of Donor Blood
 Segment closest toSegment closest to
unit is hemolyzed.unit is hemolyzed.
 May indicate bacterialMay indicate bacterial
contaminationcontamination

60. Donor Blood Inspection and Disposition
 If a unit's appearance looks questionable do the
following:
– Quarantine unit until disposition is decided.
– Gently mix, allow to settle and observe appearance.
 If bacterial contamination is suspected the unit should be
cultured and a gram stain performed.
 Positive blood cultures usually indicative of:
– Inadequate donor arm preparation
– Improper pooling technique
– Health of donor - bacteremia in donor
 If one component is contaminated, other components
prepared from the same donor unit may be
contaminated.

61. Inspection of Donor BloodInspection of Donor Blood
 Reissuing blood cannot be done unless the following
criteria is met:
– Container closure must not have been penetrated or entered in
any manner.
– Most facilities set 30" time limit for accepting units back, warming
above 6-10C even with subsequent cooling increases RBC
metabolism producing hemolysis and permitting bacterial
growth.
– Blood must have been kept at the appropriate temperature.
– One sealed segment must remain attached to container.
– Records must indicate that blood has been reissued and
inspected prior to reissue.

62. Transportation of Blood and Blood
Components
 WB and RBC
– Sturdy well insulated cardboard and/or
styrofoam container, wet ice in ziplock bag to
cool, temperature must be monitored.
– Mobile collection units should transport blood
ASAP and leave at RT if platelets are to be
made.
– In-house transport place in cooler with wet
ice and thermometer, monitor temperature
every 30 minutes.

63. Safe-T-Vue Temperature MonitorSafe-T-Vue Temperature Monitor

64. Transportation of Blood and Blood
Components
 Frozen components
– Temperature must be maintained at or below required
storage temperature.
– Use dry ice in well insulated container.
 Platelets and granulocytes
– Maintain at 20-24 C.
– Transport in well insulated containers without ice.
 Commercial coolers available to maintain at 20-
24C.

65. Transportation of Blood and Blood
Components
 Handling donor units
– Should not remain at RT unnecessarily, when
blood is issued it should be transfused as
soon as possible.
– When numerous units are removed from
fridge, remove fluid filled container with a
thermometer at same time as blood, when
temperature reaches 6 C return to fridge.

66. Records
 Must be made concurrently with each step
of component preparation, being as
detailed as possible for clear
understanding.
 Must be legible and indelible.
 Must include dates of various steps and
person responsible.

68. EXAM 1 ONLINE