Blood component processing &

some technical points about blood component

1. Blood component preparation & Preservation Bipul Chandra Sarkar Medical technologist Transfusion Medicine

2. Introduction Whole Blood: One unit of donor blood collected in suitable anticoagulant preservative solution and which contain blood cells and plasma. Blood Components: A constituent separated from whole blood, by differential centrifugation of one donor or by apheresis.

3. Components of blood

4. Components of blood

5. Whole Blood Red Cells Granulocytes Plasma Platelets FFP Fractionated products C ryoprecipitate Cryosupernatant F VIIa * F VIII* F IX* Albumin Immune Globulin PRP PLT Conc

6. Whole blood

7. Whole Blood Volume: 450ml blood. Anticoagulant: 63ml Storage temp : 4˚C (Range is 2-6˚C ) Hct- 38-44%. No component have been removed. Shelf life (it is the time of storage after which, after transfusion, at least 70% of transfused cells remain in recipient circulation) Citrate-Phosphate-Dextrose (CPD) - 28 days. CPDA-1 (adenine) - 35 days.

8. Packed red cell

9. Packed red cell Principle: RBCs are obtained by removal of supernatant plasma from centrifuged whole blood. Preparation: Centrifuge whole blood unit in refrigerated centrifuge containing the parameters RPM-3850,Time-5 Min, Temp- 4˚C Express the supernatant plasma with the help of plasma expresser.

10. Continue.. Double seal the tubing between the primary and satellite bag . Check that the satellite bag has the same donor number as that on the primary bag and cut the tubing between the two seal.

11. Packed red Cell Check that the satellite bag has the same donor number as that on the primary bag and cut the tubing between the two seal Double seal the tubing between the primary and satellite bag . Express the supernatant plasma with the help of plasma expresser . Centrifuge(RPM-3850,Time-5 Min, Temp- 4˚C) W hole blood

12. Continue.. Volume : 250- 300 ml. Storage temp : 4˚C (Range is 2-6˚C) Shelf life: (If CPDA1 anticoagulant) -35 days CONTENTS: Red cells- 65-80% Plasma – 20-35 % Some platelets, white cells storage lesion by products and anticoagulant preservative solutions . QC Requirements: PCV 80% (Range is 65-80%)

13. Platelet Rich Plasma(PRP) It is prepared from the whole blood within six-hours of collection, preferably stored at room temperature of 20-24°C .

14. Steps The blood is collected in-CPDA-1 double or triple bag. Centrifuge whole blood unit in refrigerated centrifuge containing the parameters RPM-1750,Time-11 Min , Temp- 20-24˚C. After centrifugation, the bags are taken out from centrifuge chamber with minimal disturbance and the PRP is expressed from the primary bag into the satellite bag with the help of a plasma expresser. After placing proper knots, labeling the blood group and ensuring the screening status the satellite bag is detached from the primary bag.

15. Continue.. Volume: 250-300 ml Storage temp : 22-24°C Shelf life: Maximum 48-72 hours in a platelet incubator with constant agitation. Composition: P latelet-derived growth factor Transforming growth factor beta F ibroblast growth factor I nsulin-like growth factor 1 & 2 V ascular endothelial growth factor E pidermal growth factor Interleukin 8 K eratinocyte growth factor C onnective tissue growth factor

16. Continue.. QC Requirements: As a part of internal quality control 1% of the random components units over a month are tested for pH and yield. For PRP the pH should always be > 6.2 and yield 4.5 x 10¹°/ bag ( Drugs and Cosmetics Act) and 5.5 x 10¹ °/ bag ( AABB Technical Manual). One bag of PRP generally raises the platelet count in the recipient by 5000-10000 / μl .

17. Thank you

18. Blood component processing & preservation Manik chandra shill Medical technologist Transfusion Medicine

19. Platelet Concentrate

20. Platelet Concentrate This supplies the same amount of platelets as PRP, but in lesser volume (40 - 50ml ). Principle : Platelets are harvested from whole blood following ‘light spin 'centrifugation. The platelets are concentrated by 'heavy spin 'centrifugation with subsequent removal of supernatant plasma.

21. P reparation .. Blood is collected in the triple bag system only. PRP is prepared by following the above mentioned steps. After detaching the satellite bags from the primary bag, the PRP in again spin at 4000 - 5000 rpm (high spin) for 4 to 5 minutes ,Temp :20°C - 24°C A platelet button is formed at bottom and platelet poor plasma is expressed from the 1st satellite bag to the 2nd satellite bags, leaving behind 40 - 50 ml plasma for platelet button suspension.

22. PRP is again spin at 4000 - 5000 rpm (high spin) for 4 to 5 minutes. Platelet poor plasma is expressed from the 1st satellite bag to the 2nd satellite bags, leaving behind 40 - 50 ml plasma for platelet . Platelet Rich Plasma(PRP) The blood is collected in-CPDA-1 triple bag Check that the satellite bag has the same donor number as that on the primary bag and cut the tubing between the two seal Centrifuge (1750 rpm for 11 min 20 -24°C) The bags are taken out from centrifuge chamber with minimal disturbance PRP is expressed from the primary bag into the satellite bag with the help of a plasma expresser

23. Continue.. Volume: 30 to 50 ml. Storage temp : 20°C - 24°C Shelf life: Maximum 5 days with continuous incubation & agitator. : 24 hrs if no storage cabinet. Contents : Platelet - 5.5 x 10¹° /bag Plasma - 30 to 50 ml and some white cells Q.C. Requirements: To be prepared within 8 hrs after collection, pH should be 6.2 or more at the end of storage time. Platelet count > 5.5 x 10¹° /unit.

24. Fresh Frozen Plasma

25. Fresh Frozen Plasma Plasma along with anticoagulant preservative. Prepared from blood within 8 hrs of donation. Maximum level of labile and non-labile clotting factors (about 1 IU per ml) V & VIII, proteins C and S, complement , and immunoglobulin. Good for 24 hours post thaw. Then it can be stored for 5 days as liquid plasma (labile factors V and VIII decreased).

26. Continue.. Principle : Plasma is separated from cellular blood elements and frozen to preserve the activity of labile coagulation factors. Steps Blood is collected in a CPDA-1 double or triple bag system. Centrifuge whole blood unit in refrigerated centrifuge containing the parameters RPM-3850,Time-5 Min, Temp- 4-6˚C.

27. Continue .. After the centrifuge stops, the blood bags are taken out with minimal disturbance and supernatant plasma is expressed into the satellite bag, with the help of a plasma expressor, leaving behind 80-90ml . After labeling the group and ensuring screening status, the plasma is stored at -20°C or below. The storage shelf life is one year at -20°C and 7 years at - 70 to-80°C

28. Continue .. Volume: 200-250 ml Storage temp: -20°C to -65˚C Shelf life: One year to 7 year. Contents: Per bag Factor VIII - 0.6 IU / ml Factor IX - 0.9 IU / ml Fibrinogen - 250-300 mg / bag Proteins - Albumin, globulin, etc. Q. C, Requirements: The entire process of preparation and freezing should be completed within 8 hrs after collection.

29. Continue .. 1 IU / kg of factor VIII or factor IX raises the factor VIII levels in plasma by 2% and factor IX levels by 1% respectively.

30. Cryoprecipitate Cryoprecipitate is the cold – insoluble portion of plasma that precipitates when FFP is thawed between 1-6°C

31. Cryoprecipitate contains precipitated proteins of plasma, rich in factor VIII and fibrinogen, obtained from FFP prepared within 6-8 hours of collection, subsequent thawing at 4- 6°C and removal of supernatant. Also the advantage of Cryoprecipitate is that we can administer large amount of factor VIII without overloading the recipient, especially in pediatric patients.

32. Principle: Coagulation factor VIII can be concentrated by cryoprecipitation of freshly collected plasma. Cryoprecipitation is accomplished by rapid freezing of plasma and slow thawing at low temperature . Steps:

33. Continue .. Volume: 10 to 20'ml Storage temp : Frozen -18°C or less Shelf life: Frozen - 1 year Thawed - 6 hours Contents: Factor VIII - 80 to 150 units/bag Fibrinogen - 150 to 250 mg/bag Factor Xlll - 20 - 30% of whole blood von Willebrand factor - 40-70% of whole blood. Q. C. Requirements: Thaw at 37°C Factor VIII -80 units/bag

34. Question & Answer