Blood Components 101

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

Resources

• www.aabb.org

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