1. Goals Of Blood Collection
Maintain viability and function
Prevent physical changes
Minimize bacterial contamination

2. Anticoagulants Preservative Solutions
Anticoagulants prevent blood clotting
Preservatives provide nutrients for cells
Heparin
Rarely if ever used anymore
Anticoagulant ONLY
Transfuse within 48 hours, preferably 8

3. Anticoagulants CPD CPD-A1 Storage time 21 days 35 days Temperature
Slows glycolytic activity
Adenine
None
Substrate for ATP synthesis
Volume
450 +/- 10%
Dextrose
Supports ATP generation by glycolytic pathway
Citrate
Prevents coagulation by binding calcium

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

5. Additive Solution
Add additive solution to RBCs, ADSOL, which consists of:
Saline
Adenine
Glucose
Mannitol
Extends storage to 42 days
Final hematocrit approximately 66%

6. Changes Occur During Storage
Shelf life = expiration date
At end of expiration must have 75% recovery
At least 75% of transfused cells remain in circulation 24 hours AFTER transfusion

7. Storage Lesion Biochemical changes which occur at 1-6C
Affects oxygen dissociation curve, increased affinity of hemoglobin for oxygen.
Low 2,3-DPG, increased O2 affinity, less O2 released.
pH drops causes 2,3-DPG levels to fall
Once transfused RBCs regenerate ATP and 2,3-DPG
Few functional platelets present
Viable (living) RBCs decrease

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

9. Storage Lesion Significant for infants and massive transfusion.
Other biochemical changes
ATP decreases
Potassium increases
Sodium decreases
Plasma hemoglobin increases

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

11. Preparation of Components
Centrifuge – light spin, platelets suspended
Remove platelet rich plasma (PRP)
Centrifuge PRP heavy spin
Remove platelet poor plasma
Freeze plasma solid within 8 hours
Thaw plasma at 1-4C – precipitate forms
Centrifuge, express plasma leaving cryoprecipitate. Store both at -18C
RBCs – CPD – 21 days, ADSOL – 42 days – 1-6C

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

14. Blood Component General Information
Transfusion practice
Transfusion requires doctor’s prescription
All components MUST be administered through a filter
Infuse quickly, within 4 hours
D (Rh) neg require D neg cellular products
ABO identical preferred, ABO compatible OK
“Universal donor” – RBCs group O, plasma AB

15. Blood Component General Information
Fresh Whole Blood
Candidates
Newborns needing exchange transfusion
Patients requiring leukoreduced products

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

17. Blood Components Cellular Plasma Red blood cell products Platelets
Granulocytes
Plasma
FFP
Cryoprecipitate

18. Products With Red Cells

19. Changes in Stored Blood

20. Red Blood Cells, Packed (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.

21. Leukocyte Reduced Red Cells (LR-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.

22. Leukocyte Reduction

23. 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.

24. Cell Washer to Prepare Washed Cells

25. Frozen Blood

26. 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.

27. 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

28. Platelet Products

29. 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,.
Store at 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.

30. Preparation of platelet concentrate
Plasma
RBCs
PRP
Platelet concentrate

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

32. Pooling Platelets 6-10 units transferred into one bag
Expiration = 4 hours

33. One pheresed unit is equivalent to 6-8 RD-PC.
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 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

34. Apheresis

35. Apheresis

36. Platelets Pheresis One bag (unit) from one donor
One unit is a therapeutic dose
Volume approximately 250 ccs

37. Granulocytes
Lymphocyte
Monocyte
Neutrophils
Eosinophils
Basophils

38. 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 C.

39. Plasma Components

40. Fresh Frozen Plasma – Volume 200-250cc

41. 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

42. Fresh Frozen Plasma (FFP)
Storage temperature
frozen -18 C, preferably -30 C or lower
thawed 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

43. 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

44. 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

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

46. 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

47. Cryoprecipitate – volume 15ccs

48. Irradiation of Blood Components

49. 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

50. 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.

51. Irradiation of Blood Components

52. Inspection of Donor Blood
Segment closest to unit is hemolyzed.
May indicate bacterial contamination

53. Safe-T-Vue Temperature Monitor

54. 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 C.
Transport in well insulated containers without ice.
Commercial coolers available to maintain at 20-24C.

55. Cont… Blood processing:- Donor selection Blood collection
Component preparation
Storage , issue and transportation
Contract testing

56. Donor selection
Principle of self-sufficiency from voluntary and non- remunerated donations have been recommended and promote by the Council of the Europe Recommendations No.R(95)14
Main purpose is to determine whether the person in good health, in order to protect the donor against damage to his/her own health, and to protect the recipient against transmission of disease or drugs which could be detrimental to the patient.

57. Blood collection & processing
Aseptic technique
Seal closed method
Immediate storage at 1-6ºC
Components preparation has to be done within 6 hours after collection
Labels/records : ABO and Rh grouping
Screening, expiratory date and volume of the blood

58. QC of blood component preparation
Whole blood:
Frequency of control: 1% of all units with minimum of 4 units per month
Storage :- 2ºC to 6 ºC, for CPDA-1 the storage time is 35 days, CPD & CD2D – 22days.
On expire date :- measure HCT, pH, total Hb , K+ and perform sterility assays

59. QA:-
Volume : 450ml ± 10 % of body volume excluding anticoagulant
HCT : 40±5%
pH > 6.5
K < 27mmol/L
Hb minimum 45g/unit
Sterility : no growth

60. Red cell concentrates
Perform the same assay as for Whole blood on the expiry date
Storage : 2-6º C, for 35 days if prepared from WB collected in CPDA-1
QA:
Volume : 280ml± 50ml, frequency of control 1% of all units
HCT :
pH > 6.5
K < 78 mmol/L
Hb : minimum 45g/unit
Sterility : no growth

61. Platelet concentrates:
Prepared within 6H of blood collection
Must evaluate at least 4 platelet preparations monthly for platelet count,pH and plasma volume
Platelets should be selected from each centrifuge in use
The Tº at which pH is measured should be the same as stored
Label the volume,
Storage : 20-24ºC
Tº should be recorded at least every 4H during storage.

62. QC
Volume > 40ml
pH :
Plt count : at least 5.5 x 1010 /bag in at least 75% of the units tested at the end of the storage.By apheresis : minimum 3 x 1011/bag platelets in at least 75% units tested
WBC contamination: < 2 x 103/bag
RBC contamination: < 2 x 109/bag
Macroscopic appearance : no visible platelets aggregates
Sterility : no growth

63. Fresh Frozen Plasma Every 10 unit/week estimate the volume Storage:
24 months at below –30ºC
12 months at –25 to –30ºC
3 months at –18 to –25ºC
Thawed at Tº between 30-37ºC and transfused within 24H after thawing QC
Volume: ml
Factor VIIIc : > 0.7IU/ml- every 2 months
No leakage after pressure in plasma extractor, before freezing and after thawing

64. QC Macroscopic : no abnormal color or visible clots Residual cell:
Red cell: < 6.0 x 109/l
Leukocyte: < 0.1 x 109/l
Platelets : < 50 x 109/l

65. Cryoprecipitate Assayed on at least 4 bags/ month –for factor VIII
Storage:
24 months at below –30ºC
12 months at –25 to –30ºC
3 months at –18 to –25ºC
Must be thawed at 37ºC and used within 6H

66. QC Volume : 10-20 ml Factor VIII : > 70 IU/unit
Fibrinogen : > 140 mg per unit
Macroscopic : homogenous
Sterility: no growth

67. Granulocytes Prepared by apheresis Storage: 20-24ºC for 24H QC
Volume : < 500ml
Granulocytes : > 10 x 109 per unit- present in at least 75% of all units tested

68. Transportation
A system must be use to ensure that all blood and blood components shipped by or received into a blood bank or blood transfusion service have been maintained within T required.
All liquid RBC components kept at T of 1-10ºC during transport
All component routinely stored at 20-24ºC should be maintain T during shipment
All frozen components should be transport in frozen state at –18ºC or colder
Periodic T check and documented to ensure the transportation adequate to meet the criteria

69. QC FOR EQUIPMENT

71. Quality control for reagents
Select the reagent with high specifications- reference preparation has been established for ABO, Rh and anti-human globulin (AHG) by FDA
Color codes by the FDA:
Blue for anti-A
Yellow for anti-B
Green for AHG
Use according to manufacturer's instruction
The new reagent has to be assessed & confirmed satisfactory
The appearance each reagent has to be checked each day
The reactivity and specificity has to checked each new lot