blood components 101
TRANSCRIPT
Blood Components 101
Kim-Anh Nguyen, MD, PhDTransfusion Medicine Symposium
August 6, 2016
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
Objectives
• Describe the acceptable parameters for a therapeutic dose of red cells.
• Describe the acceptable parameters for a therapeutic dose of platelets.
Component Overview
• Red cells• Platelets• Plasma• Cryoprecipitate AHF• Modified components
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
Whole Blood → RBC
www.nature.com
Anticoagulant-Preservative Solutions
• Citrate
• Dextrose
• Adenine
• Na phosphate
• Prevents coagulation
• Glycolytic pathway
• ATP synthesis
• Buffer
Anticoagulant - Preservatives
CPD CP2D ACD-AApheresis 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)
Additive Solution (AS)
• Extends shelf life of RBCs → 42 days• Replaces plasma in RBCs
– More volume in Frozen Plasma unit• Hct 55-65%
RBC Additive Solutions
AS-1 (Adsol)
AS-3(Nutricel
)
AS-5(Optisol)
AS-7(SOLX)
Dextrose 2200 1100 900 1585Adenine 27 30 30 27NaH2PO4 0 276 0 0Na2HPO4 0 0 0 170Mannitol 750 0 525 1000Na Bicarbonate
0 0 0 218
Na Chloride 900 410 877 0Na Citrate 0 588 0 0Citric Acid 0 42 0 0
(100/450mL bag or 110/500mL bag)
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
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
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
Frozen Plasma
• Store at -20C• Shelf-life 1 year• Thawing Methods (30-37C)
– Waterbath– Microwave– Ultrasound
Thawed Plasma
• “FFP & FP24, thawed”– 1-6C– 24 hours (no FDA variance needed)
• “Thawed plasma”– 1-6C– 5 days– closed system only
Thawed PlasmaClotting Factor Changes Over Time
AABB Technical Manual, 17th ed. 2011Downes et al. Transfusion (2001) 41:570.
Cryoprecipitate AHF
• 5-20mL Volume
• Fibrinogen ≥ 150 mg / unit• FVIII ≥ 80 IU / unit• vWF, FXIII, fibronectin
Cryo Pooling & Storage
• May be pooled during production– 5U per pool
• Store at ≤ -18 ºC • Shelf-life 1 year
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
WB Derived Platelets
www.researchgate.net
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
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
Pheresis vs. WB platelets
Apheresis WB Volume 250-400 mL 40-70 mLPlatelet 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 daysStorage 20-24C + agitation 20-24C + agitation
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
Pediatric transfer bags
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
DIN
• Longer : W 1234 06 123456
• Country: W• Facility: 1234• Year: 06• Unique ID: 123456 ( 100 years)
Blood Component Label
4 quadrants, black & white onlyBarcode readable elements• Donor Identification Number (DIN) • Product code • ABO/Rh Type includes donation type • Expiration Date and time• Special testing performed
Blood Component Label
Component Modifications
• Leukoreduction• Washing• Irradiation• Freezing
Leukocytes in Blood
Whole blood 109
Red blood cells (RBC) 108 - 109
Whole-blood platelets 107 - 108
Apheresis platelets 106 - 108
FFP < 104
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
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%)
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
Washed Components
• Red cells– 1-6C– 24 hours
• Platelets– 20-24C + agitation– 4 hours
Frozen Red Cells
• Freeze in 20-40% glycerol• Storage
– Store at -65C– Shelf-life10 years
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
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
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
Irradiated components
Shelf-life• Red cells (shorter of the following)
– 28 days – Original date
• Platelets: Original date
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
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
Acceptable parameters - Platelets
Pheresis• 250-400 mL volume• >3.0 x 1011 platelets/Unit• <5 x 106 WBC/Unit• ~1 x 109 platelets/mL
