This document summarizes blood component production and acceptable parameters. Red blood cells can be collected in CPD, CPDA-1, or additive solutions with a volume of 225-400mL and hematocrit of 55-65%/65-80% with a shelf life of 21-42 days. Platelets can be collected through apheresis with a volume of 250-400mL, platelet count of >3.0 x 1011 per unit, and <5 x 106 white blood cells per unit. The document also reviews definitions, anticoagulants, storage conditions, and quality control parameters for blood components.

1. Blood Components 101
Kim-Anh Nguyen, MD, PhD
Transfusion Medicine Symposium
August 6, 2016

2. Disclosures
• I have no financial relationships
related to this presentation.
• I will not be speaking about any
specific commercial product,
device, or medication.
• I will not be speaking of any off-
label use of medications or
devices

3. Objectives
• Describe the acceptable
parameters for a
therapeutic dose of red
cells.
• Describe the acceptable
parameters for a
therapeutic dose of
platelets.

4. Component Overview
• Red cells
• Platelets
• Plasma
• Cryoprecipitate AHF
• Modified components

5. Definitions
Differential centrifugation separate components
along density gradient
• Light (Soft) Spin
– 2000 RPM for 3 min
– RBCs & Platelet-rich plasma (PRP)
• Heavy (Hard) Spin
– 5000 RPM for 5-7 min
– RBCs-WBC-Plts & Platelet-poor plasma
– Platelets & Platelet-poor plasma

6. Whole Blood → RBC
www.nature.com

7. Anticoagulant-Preservative
Solutions
• Citrate
• Dextrose
• Adenine
• Na phosphate
• Prevents coagulation
• Glycolytic pathway
•  ATP synthesis
• Buffer

8. Anticoagulant - Preservatives
CPD CP2D
ACD-A
Apheresis
CPDA-1
Trisodium Citrate 26.3g/L 26.3g/L 22.3g/L 26.3g/L
Citric Acid 3.27 g/L 3.27 g/L 8.0 g/L 3.27 g/L
NaH2PO4 2.22g/L 2.22g/L 0 2.22g/L
Dextrose 25.5 g/L 51.1 g/L 24.5 g/L 31.9g/L
Adenine 0 0 0 0.275g/L
Shelf Life 21 days 21 days 21 days 35 days
*Apheresis Only
(63-70 mL/bag or 14mL/100mL of WB collected)

9. Additive Solution (AS)
• Extends shelf life of RBCs → 42 days
• Replaces plasma in RBCs
– More volume in Frozen Plasma unit
• Hct 55-65%

10. RBC Additive Solutions
AS-1
(Adsol)
AS-3
(Nutricel)
AS-5
(Optisol)
AS-7
(SOLX)
Dextrose 2200 1100 900 1585
Adenine 27 30 30 27
NaH2PO4 0 276 0 0
Na2HPO4 0 0 0 170
Mannitol 750 0 525 1000
Na Bicarbonate 0 0 0 218
Na Chloride 900 410 877 0
Na Citrate 0 588 0 0
Citric Acid 0 42 0 0
(100/450mL bag or 110/500mL bag)

11. CPDA-1 vs. AS Red Cells
CPDA-1 Red Cells
• 225-350mL total volume
• 160-275mL RBCs
• 50-80g Hgb
• 65-80% Hct
• 35 day shelf-life
AS Red Cells
• 300-400mL total volume
• 160-275mL RBCs
• 50-80g Hgb
– ~60g Hgb apheresis RBC
• 55-65% Hct
• 42 day shelf-life

12. Frozen Plasma
• Processed from WB
• Collected by apheresis
– Concurrent: Part of RBC or platelet
procedure
– “Plasmapheresis”: plasma only procedure
• Volume 200-800mL
– WB derived ~200-250mL
– Pheresis variable, divided or “Jumbo” units

13. Plasma Components
• Fresh Frozen Plasma (FFP): 6-8 hrs of
phlebotomy
• Frozen Plasma (FP24):24 hrs of
phlebotomy
• Freezing methods
– Dry ice-ethanol bath
– Blast freezer (-18C)
– Mechanical freezer (-65C)
• Equivalent therapeutic indications

14. Frozen Plasma
• Store at -20C
• Shelf-life 1 year
• Thawing Methods (30-37C)
– Waterbath
– Microwave
– Ultrasound

15. Thawed Plasma
• “FFP & FP24, thawed”
– 1-6C
– 24 hours (no FDA variance needed)
• “Thawed plasma”
– 1-6C
– 5 days
– closed system only

16. Thawed Plasma
Clotting Factor Changes Over Time
AABB Technical Manual, 17th ed. 2011
Downes et al. Transfusion (2001) 41:570.

17. Cryoprecipitate AHF
• 5-20mL Volume
• Fibrinogen ≥ 150 mg / unit
• FVIII ≥ 80 IU / unit
• vWF, FXIII, fibronectin

18. Cryo Pooling & Storage
• May be pooled during production
– 5U per pool
• Store at ≤ -18 ºC
• Shelf-life 1 year

19. Thawed Cryoppt
• Thaw at 30-37 C
• May be pooled post-thaw
– 10U per adult dose pool
• Store at Room Temperature
• Thawed cryoppt shelf-life:
– 4 hrs Open system
– 6 hrs Closed system
• ABO Compatibility not required for transfusion

20. WB Derived Platelets
www.researchgate.net

21. Apheresis Platelets
• Automated Collection Device
– Trima (Terumo)
– Amicus (Fenwal)
• Single or double needle
• Leukoreduction occurs during
procedure
• 50-120 minutes
• 1-4 Units (doses) / procedure

22. Platelet Storage
• Blood must be received at
– ≥ 20 ºC (≥ 68 ºF)
• Storage:
– 20-24 ºC (68-75 ºF)
– Continuous gentle
agitation
– Shelf-life 5 days

23. Pheresis vs. WB platelets
Apheresis WB
Volume 250-400 mL 40-70 mL
Platelet Count >3.0 x 1011 >5.5 x 1010
Platelet
Concentration
~1 x 109/mL ~1 x 109/mL
Adult Dose 5-10mL/kg;
1U
5-10mL/kg;
4-6U
Shelf-life 5 days 5 days
Storage 20-24C + agitation 20-24C + agitation

24. Platelet Tx for infants
• Pheresis platelets or WB platelets
• 1U pheresis = 4-6 U WB platelets
• Dose same way
• 1U pheresis may be “split” into several aliquots
for multiple dosing
– Done in blood bank by request
– Reduces donor exposure
– Aliquots may be given to other patients

25. Pediatric transfer bags

26. ISBT128 Label
Global standard for identification, labeling and
information processing of human blood, tissue and
organ products across international borders and
disparate health care systems
• Unique donation identification # (DIN)
• Blood collection center listed
• Product Code is expandable
• Adopted in USA in 2008

27. DIN
• Longer : W 1234 06 123456
• Country: W
• Facility: 1234
• Year: 06
• Unique ID: 123456 ( 100 years)

28. Blood Component Label
4 quadrants, black & white only
Barcode readable elements
• Donor Identification Number (DIN)
• Product code
• ABO/Rh Type includes donation type
• Expiration Date and time
• Special testing performed

29. Blood Component Label

30. Component Modifications
• Leukoreduction
• Washing
• Irradiation
• Freezing

31. Leukocytes in Blood
Whole blood 109
Red blood cells (RBC) 108 - 109
Whole-blood platelets 107 - 108
Apheresis platelets 106 - 108
FFP < 104

32. Filter Leukoreduction
• Prestorage (donor center)
– 3-5 days post collection
• Bedside (hospital)
• Pre-storage LR preferred
– > reliable
– < variable
• 100% of RBCs and platelets at BBH are pre-
storage LR

33. Leukoreduction QC
• Red cells
– ≤ 5 x 106 WBC/unit (≥ 95%)
– ≥ 85% of original red cells
• Apheresis platelets
– ≤ 5 x 106 WBC/unit (≥ 95%)
• WBD platelets
– ≤ 8.3 x 105 WBC/unit (≥ 95%)

34. Washing Components
• Saline (1-2 L) removes
– ≈ 99% of plasma
– Potassium
– Anticoagulant
– Additive solution
– Cellular debris
• Product loss
– 10-20% red cells
– 20-30% platelets

35. Washed Components
• Red cells
– 1-6C
– 24 hours
• Platelets
– 20-24C + agitation
– 4 hours

36. Frozen Red Cells
• Freeze in 20-40% glycerol
• Storage
– Store at -65C
– Shelf-life10 years

37. Thawing & Deglycerolyzing
• Thaw 20-25 min in waterbath or dry warmer
• Wash with solutions of ↓osmolality
– Cobe 2991 – open system
– Haemonetics ACP215 – closed system
• QC: >80% recovery
• Shelf-life of deglyc’d RBCs at 1-6C
– 24hrs open system
– 14 days closed system

38. Irradiation
• Objective: inactivate T-lymphocytes
• Prevent Graft vs. host disease (TA-GVHD)
• Red cell damage
– Decreased % recovery after transfusion
– Increased potassium levels in unit (2x)
• No damage to platelets

39. Irradiation
• Methods:
– Cobalt-60
– Cesium-137
– X-ray
• Dosage:
– Minimum 25 Gy (2500cGy) to center, 15 Gy
to any portion
– Maximum 50 Gy to center
• Method to ensure irradiation has occurred

40. Irradiated components
Shelf-life
• Red cells (shorter of the following)
–28 days
–Original date
• Platelets: Original date

41. Summary
• Blood component production (collection,
processing) has evolved
• Variety of volumes, storage media,
specifications
• ISBT128 label → extensive information
• Production uses validated methods
• Components undergo routine QC

42. Acceptable parameters -
RBC
• CPD, CPDA-1, AS-1, AS-3,
AS-5
• 225-400 mL Volume
• 55-65% / 65-80% Hct
• Shelf-life 21-42 days
• LR: < 5 x 106 WBC/Unit

43. Acceptable parameters -
Platelets
Pheresis
• 250-400 mL volume
• >3.0 x 1011 platelets/Unit
• <5 x 106 WBC/Unit
• ~1 x 109 platelets/mL

44. Resources
• www.aabb.org
• www.bbh.org: Circular of Information
• knguyen@bbh.org; 808-848-4705