1. Blood Component Preparation and Therapeutic Utilization
Prof. A. Pourazar
Immunohematologist and Transfusion medicine
School of Medicine, Isfahan Medical Science University
IRAN

2. Whole Blood
Red Blood cells, Platelets and White Blood Cells- all suspended in Plasma constitute Whole Blood.

3. Whole Blood Unit
After centrifugation whole blood separates into the plasma and platelets on top and packed red blood cells on the bottom.
A plasma expresser is used to squeeze the plasma and pla
telets off the top and leave only the red blood cells in the original bag.

4. Blood Components Random donor Platelets Packed Red Blood Cells
Plasma Product
When whole blood is separated into it’s component parts we now have Blood Components!!

5. Labeling the Bag
Base Label: Bag comes with label indicating manufacturer and anticoagulant/ preservative.
2. Completed label: Paste ABO Group, Rh Type, Collection date, collecting facility etc. on label
See Harmening p

6. Base label on bag at time of collection.
Blood Bag label when testing is complete.

7. RBC Components

8. Packed RBCs Approx. half the volume of Whole Blood
Same RBC mass therefore same oxygen carrying capacity
Total Volume: ml
Hematocrit <80% for CPDA-1 anticoagulant/ preservative.
Expiration Date: depends on sterility and anticoagulant/preservative
CPDA days closed system
AS days closed system
Open System - 24 hours

9. RBC:Anticoagulant/Preservative Solutions
Purpose of RBC Preservation
Designed to prevent clotting and maintain red cell viability and function during storage.
Anticoagulant-Preservative Contents
Citrate: anticoagulant (chelates calcium)
Dextrose: ATP generation via glycolytic pathway
Adenine: Acts as a substrate for RBC synthesis of ATP
Sodium diphosphate: Buffer to control decrease of pH expected from generation of lactic acid over time.

10. RBC Anticoagulant/Preservative Solutions
Cold Temperature
RBC components are kept at 1-6oC in a monitored refrigerator
Inhibits bacterial growth
Slows glycolytic activity: But RBCs continue metabolic activity during storage consuming nutrients and depleting intracellular energy sources.

11. RBC Component Storage Lesion
Definition: The biochemical changes that occur during storage of RBC components.
Changes in Red Blood Cells include:
 2,3-DPG,  K+,  ATP  Na+
Changes in the PLASMA include:
 K+,  Ammonia,  pH, and  haemolysis

12. Indications for Transfusion of Packed Red Blood Cells
Need for blood depends on patients:
Oxygen demand
Cardiac Output
Haemoglobin concentration (Hgb and Hct)
Each unit of transfused RBCs increases
-Hemoglobin by 1.0 g/dL
-Hct by 3%.

13. Indications for Transfusion of Packed Red Blood Cells…
RBC Transfusion Categories
Iatrogenic: neonatal transfusions often result from withdrawal of too much blood for testing.
Decreased Bone Marrow Production: Leukemia, aplastic anemia, etc.
Decreased RBC Survival: Hemolytic anemia, etc.
Surgical/Trauma: Excessive bleeding

14. Indications for Transfusion of Packed Red Blood Cells…
Fresh Red Blood Cells (<7 days old)
Neonatal transfusion. Why?
Massive Transfusion is defined as replacing the patients entire blood volume within 24 hours.
Why would we want to transfuse fresh blood in this situation?
2,3-DPG levels drop within 10 to 15 days of collection to levels that will NOT adequately oxygenate the tissues.
Also want to avoid transfusion of excess K+, ammonia and haemolysis.

15. RBC Components continued…
Leukocyte reduced RBC: to a level of <5.0 X 106 per unit
Leukocyte filtration: removes WBCs, see p242
Washing- removes WBCs, not efficiently, 24 hour outdate, -see p242
Freezing/Deglycerolization: Frozen RBC unit is thawed and washed to remove both glycerol and WBCs, see p242

16. Indications for Transfusion of Leukocyte Reduced RBCs
Febrile non haemolytic transfusion reactions
Patient history of febrile transfusion reactions
HLA allo-immunization: antibody to HLA antigens
Transfusion related acute lung injury (TRALI)
Transfusion associated Graft vs. Host disease (TA-GvHD).
Transfusion related immune suppression:
- Reduces risk of transmission of CMV and HIV.

17. RBC Components continued…
Frozen, Deglycerolized RBC
Freezing RBCs in glycerol gives minimal damage to the cell.
>10 years Shelf life, while in frozen, for ‘rare’ blood types
Free (almost) of WBCs, Platelets & Plasma
Indicated for patient with anti-IgA;
Intra-uterine transfusion (as lacks WBCs)
Two methods: Page

18. Platelet Components
Includes Random donor platelets, Single donor platelets and Pooled Platelets.

19. Platelet Components Preparation
Random Donor From a Whole Blood (WB) Unit at kept 20-24oC,
Step 1: WB centrifuged using a Soft (light) Spin for 2-3 min at 3200 rpm (to keep the platelets in the plasma).
Step 2: Separate platelet rich plasma from RBCs and using a Hard (heavy) Spin (5 min at 3600 rpm) to settle platelets.
Step 3: Express off all but ml of plasma and let platelets rest and resuspend at RT for 1-2 hours
Single Donor (by apheresis)

20. Platelet Components Random Donor Platelet
At least 5.5 x 1010 platelets/unit
Store at 20 to 24oC with continuous agitation
Suspended in ml plasma
Shelf life of 5 days
Single Donor Platelet - Apheresis
At least 3.0 x 1011 platelets/unit
Store at 22 to 24oC with agitation
Suspended in 300 ml plasma (equivalent to 4-8 random donor platelets)

21. Platelet and Plateletpheresis
Platelet Increment (desired): Patient platelet count should increase by 5-10,000/ random donor platelet transfusion and 30-60,000/ per single donor platelet.
ABO and Rh Compatibility
Paediatric:
Transfuse ABO/Rh compatible units
Volume may be reduced by
Adults:Want to use ABO compatible with adults but it is NOT necessary. Need to be careful since there is ml of plasma per unit.
No cross match is necessary.

22. Platelet refractoriness (Poor platelet increment)
Transfusion of platelets is indicated when patient’s bleeding results from decreased platelet count. Platelet Products
Platelet refractoriness (Poor platelet increment)
ITP (idiopathic thrombocytopenic purpura) and
TTP (thrombotic thrombocytopenic purpura)
Due to antibody to platelets: don’t transfuse unless absolutely necessary (ITP and TTP). Adding fuel to the fire.
Splenomegaly, drugs or sepsis can also cause platelet refractoriness.

23. Indications for Transfusion of Platelets
Thrombocytopenia Decreased platelet count
Causes:
Decreased Platelet production: chemo-therapy, malignancy, etc.
Increased Platelet destruction: DIC
Decrease by dilution: Massive transfusion
Transfusion indicated, if associated with bleeding

24. Plasma Components Fresh Frozen Plasma
Plasma expressed from Whole Blood, frozen within 8 hours of collection for CPDA-1 anticoagulant (6 hrs for ACD).
Frozen at -18oC: 1 year expiration date
Frozen at -65oC: 7 year expiration date
Thawed: 24 hour kept at 1-6oC
ml total volume
Contains all clotting factors

25. Indications for Transfusion of Plasma Products
Fresh Frozen Plasma (FFP)
Treat multiple coagulation factor deficiencies such as DIC, liver failure, vitamin K deficiency, or massive transfusion
Good for factor deficiency where there is no suitable clotting factor concentrate: Factor XI deficiency
NEED to be ABO compatible. No cross-match needed
Contraindicated for volume expansion and protein replacement.

26. Plasma Substitutes Colloid Solutions Albumin: 5% and 25%
Plasma Protein Fraction: 5% albumin and non albumin plasma proteins
Crystalloid Solutions
Normal saline, lactated Ringers, dextran
Indications: Volume expansion/protein replacement without risk of transfusion transmitted viruses.
Advantages over plasma: No disease transmission, cost, reduced risk of allergic reactions.

27. Cryoprecipitated Antihemophilic Factor (Cryo)
Cold insoluble portion of Plasma
Preparation Procedure (Page 246)
Take FFP and thaw at 1 to 6oC until it becomes slushy (14-16 hours)
centrifuged using Hard Spin (5-7 min at 3500 rpm).
Remove cryo-poor plasma leaving about ml of plasma with Cryo-precipitates.
Freeze Cryo at -18oC for 12 months

28. Cryoprecipitated Antihemophilic Factor (Cryo)
Contains at least 80 units of AHF and mg Fibrinogen
Also contains Factor XIII and von Willebrands factor
Must be transfused within 6 hours of thawing or within 4 hours of pooling
Once thawed store at room temperature until transfusion

29. Indications for Transfusion of Cryoprecipitated AHF
Primary use: intravenous supplementation of Factor XIII and fibrinogen
Topical Use: Fibrin sealant (glue) in surgery
Factor concentrates (i.e. Factor VIII) have replaced CRYO in many situations because of reduced risk of transmission of disease.
See Table 16-4, Page 349 Harmening

30. Granulocyte Concentrates
Prepared by Pheresis (Chapter 17)
Single donor component
Should contain 1 x 1010 granulocytes/unit
ml plasma and red cells
Must be ABO and Crossmatch compatible with recipient
Store at 20oC to 24oC
Shelf life is 24 hours

31. Indications for Transfusion of Granulocyte
Patient should meet the following conditions:
Neutropenia: WBC count <500/mL
Fever for hours, positive bacterial or fungal cultures, unresponsive to antibiotic therapy.
Myeloid hypoplasia
A reasonable chance for recovery of marrow function.
Component should be irradiated and CMV seronegative.

32. Irradiated Blood Components
Gamma irradiation of component
Purpose: Inactivate donor lymphocytes to prevent Graft Vs. Host disease (GVHD). Only accepted method.
Expiration date: 28 days (RBCs) from irradiation or original outdate - which ever comes first.

33. Indications for Transfusion of Irradiated Blood Products
Congenital Immunodeficiency's: SCID, Di George Syndrome
Hodgkin's Lymphoma
Bone Marrow Transplant
Intrauterine Transfusion to Fetus
Exchange transfusion: Neonate
Transfusion of first degree relative: Blood relative

34. ALL BLOOD COMPONENTS ARE ADMINISTERED THROUGH A FILTER!
Necessary to remove any accumulated debris that may be present in the blood component such small clots, fibrin, etc.

35. Autologous Blood Transfusion
Donation from the recipient,
Rare blood type
Having antibody to high frequency antigen
Having a presence of multiple CSA
Useful in planned surgery
Large number of units by leap-frog method