BLOOD COMPONENT PREPARATION

1. BLOOD COMPONENT PREPARATION

2. This presentation will enable participants to Understand the basic principles and procedure of Component Separation Know the different components that can be prepared in a blood bank LEARNING OBJECTIVES

3. Transfusion service – certain patient goals Cater to patient needs Provide blood and components which are Safe Pure Potent Effective Policies and procedures so that goals are met INTRODUCTION

4. Development of component therapy • Earlier only whole blood • Polyvinyl Chloride (PVC) bags introduced in India in late ’80s • Treatment in various diseases/conditions Cancer : platelets DIC : plasma, platelets, cryo Haemophilia : cryo

5. Definitions Blood product Any therapeutic substance prepared from human blood Whole blood Unseparated blood containing anti-coagulant preservative Blood component A constituent of blood, separated (component from whole blood separation) eg., red cells, platelets, FFP Plasma derivative Human plasma proteins prepared (fractionation) under pharmaceutical manufacturing conditions eg., albumin

6. COMPONENT PREPARATION is “manufacturing” SOPs - outline all details of procedures Standardization Quality assurance Equipment for preparation and storage - must be maintained and reassessed CARDINAL PRINCIPLES

7. BLOOD COMPONENTS STANDARD Whole blood Packed Red Cells Platelets PRP, RDP, SDP Fresh Frozen Plasma (FFP) Cryoprecipitate SPECIALIZED Saline-washed Red Cells Frozen Red Cells Leucodepleted products Irradiated products

8. Planning a component lab Type of hospital and bed strength Trained manpower Adequate AC space (+ 50 m2) QC program Equipment License from Regulatory authorities Double, triple or quadruple bags

9. Blooddonors Fulfill criteria Hb ≥ 12.5 gm/dl Weight - 45 kg (350 ml) - 55 kg (450 ml) No Aspirin < 3 days Single bold venipuncture, free flow of blood Collection - time < 10 minutes with frequent mixing

10. Equipment for components Weighing balance Two pan balance Refrigerated centrifuge Laminar air flow bench Deep freezers (-40,-80oC) Platelet shaker/ incubator Refrigerated water bath Plasma expressor Tube sealer • Additional • Sterile tubing welder • Gamma irradiator • Cell separator (apheresis)

11. Multiple integrated blood bags

12. Quadruple Blood Bags

13. Preparation protocol Counter balancing Weighing Centrifugation Expression

14. } Packed Red Cells Centrifugation - Principle Blood cells have different Sedimentation Coefficients } Plasma } Buffy Coat

15. Protocol for preparation of Red Cells and FFP Whole Blood Soft spin at 4oC Red cellsPlasmafreeze at -30oC FFP Within 8 hours

16. Protocol for preparation of Platelets Whole blood Soft spin at 22oC within 8 hrs Red cells Platelet rich plasma (PRP) Hard spin at 22oC Plasma Platelet Conc. (RDP)

17. PRECENTRIFUGATION Allow for 2 Hours of Resting Time Maintains pH of Platelets - Better Separation CENTRIFUGATION • Proper balancing and Packing of Cups - Consistent Yield - Good Interface • Speed and Time of Centrifugation • - Dictates yields - Cellular injury • Gentle Handling - prevents mixing at interface • Resuspension of platelets before storage

18. Platelet Preparation Steps

19. Platelet Preparation Buffy Coat Method

20. Triple Bag showing separated blood components Platelets Plasma Packed Red Cells

21. Preparation of Cryoprecipitate Fresh Frozen Plasma Slow thaw at 4oC Thaw in cryobath at 4oC in Cold Room or Hard spin at 4oC Blood Bank Refrigerator Cryopoor plasma Cryoprecipitate

22. Storage and shelf life of components Component Storage Shelf Compatibility temp life Red cells + 2o-6oC 35 days ABO / Rh Red cells + 2o-6oC 42 days ABO / Rh with additive solution FFP - 30oC 1 year ABO CPP - 30oC 5 year ABO Platelets + 22oC 5 days preferably ABO match Cryoprecipitate - 30oC 1 year any group

23. Specialized equipment Sterile tubing welder Cell separator

24. Leucodepleted blood (1) • Methods • Washing (saline washed red cells) • Freezing • Buffy coat removal • Microaggregate filtration • Specific leucodepletion filters • Low leucocyte apheresis devices (cell separators)

25. Leucodepleted Blood (2) Of various methods Leucocyte filters most efficient – 99.99% (4 log depletion) Leucocyte filters can be Red cell filters or Platelet filters Leucofiltration can be done - At the bedside - In the laboratory before issue - Leucodepletion using inline filters

26. Concept of Log Reduction 1010 109 108 107 106 105 Whole Blood 1 log Reduction (90% reduction) Buffy Coat Depleted Red Cells Leucofiltered products 3 log Reduction (99.9%%)

27. IRRADIATED BLOOD For Gamma irradiation (caesium or cobalt) dose is 25 Gy Prevents transfusion associated graft-versus-host disease (TA GvHD) INDICATION : Bone Marrow Transplant Congenital immunodeficiency Premature infants Intrauterine transfusions First degree relatives

28. Plasma Derivatives They are prepared in fractionation centres from plasma pools • Albumin • Plasma Protein Fraction • Factor VIII concentrate • Fibrinogen • Immunoglobulins • Other coagulation Factors Plasma derivatives are not blood components, which are prepared in the blood bank

29. LEARNING OUTCOME At the end of this presentation participants will have understood the process involved in separation of blood components and the various blood components that can be prepared