Preparation of Blood components

Concentrated RED cells

Packed red cells are prepared by removing most of the plasma from a unit of whole blood. Red cells are prepared by removing most of the plasma from a unit of whole blood. Red cells have higher specific gravity than plasma, the red cells settle in the lower portion of the bag due to the gravitational setting (sedimentation) or centrifugation.

Red blood cells concentrate should be prepared from the whole blood collected in plastic bags, preferably in double or multiple plastic bags systems. Plasma is separated from red cells following either centrifugation or undisturbed sedimentation at any time before the expiry date of blood. If system is in use the expiry date of red cells shall be the same as whole blood. However, if an open system is used, the expiry date shall be 24 hours after separation.

Procedure :

1.    Collect appropriate volume of donor blood in CPDA double bag or triple bag.

2.      Store at 2 - 6°C till processed.

3.      Place the bags in the buckets of refrigerated centrifuge and balance the opposite bags accurately.

4.      Centrifuge at heavy spin (5000x g) for 5 minutes at 2 - 6°C

5.      Express approximately 3/4 of the plasma in to the satellite bag.

6.      Seal the tube between primary and satellite bags with plasma and separate the satellite bag.

7.      Label the product with blood group & type; Keep the red cells at 2 - 6°C and plasma at minus 30°C or below.

(2) Fresh Frozen Plasma (FFP):

FFP is plasma obtained from a single donor either by normal donation and rapidly frozen within 6-8 hours of being collected. It contains all coagulation factors and great care must be taken during collection, freezing and thawing to preserve their activity.

Collection of Blood:

1.     Blood should be collected by a clean, single venipuncture.

2.     Flow should be rapid and constant.

3.    Total time taken to collect 450 ml of blood should not be more than 8 minutes

 

 

Procedure: -

 

1.    Collect appropriate volume of blood in 350-450 ml CPDA double bags systems or 450 ml SAGM/Adsol triple bags system.

2.     Store at 4°C or in air-conditioned room till processed but not for more than 6-8 hours.

3.     Place bags in the bucket of the refrigerated centrifuge, balance them accurately and centrifuge at heavy spin (500xg for 5 ninutes) at 4°C.

4.    Express approximately four fifth of the plasma in to a satellite bag, if blood is icollected in CPDA triple/double bag system. Double seal the tube between primary bag and the satellite bag having plasma and separate them. If blood is collected in SAGM or Adsol triple bag system, express all plasma in satellite bag.

5.    Label the plasma bag and rapidly frozen for storage at -30°C or below.

Note:   FFP should be separated from the whole blood at not later than 6-8 hours of                    collection. F.F Ponce thawed it shall be used within 6 hours and in any event                         within 12 hours if kept at 2-6°C.

(3) Random donor platelet concentrate:

Platelet concentrate is prepared from 450 ml/350 ml whole blood kept at room temperature (20 - 22 °C) and within 6-8 hours of collection. Platelet concentrate is prepared by centrifugation of a single unit of whole blood collected with a smooth venepuncture and a continuous flow of blood. Platelet concentrate should be separated from whole blood by centrifugation at 22 °C + 2 using either platelet rich plasma or buffy coat method.

 Procedure:

1.     Collect blood by a clean, single venipuncture in to a triple or quadruple bag. >

2.    Keep the blood at room temperature (20-22°C) before preparing the platelets, for more than 6 hours.

 PRP method:

1.    Keep the bags in the buckets of refrigerated centrifuge and balance them accurately. Centrifuge the blood bag at 20-22°C at light spin (PRP method) for appropriate time.

2.    Separate 4/5 of the platelet rich plasma in to one satellite bag if triple bag is used. Double seal the tubing between the primary bag and the satellite bags. Separate the primary bag with red ceils, if SAGM bag is used transfer all PRP in to satellite bag and add additive solution in to red cells and separate the red cells bag.

3.   Centrifuge the bag with PRP and another satellite bag at 20-22°C at heavy spin for appropriate time.

4.    Express Supernatant plasma in to another empty satellite bag.

5.   Leave approximately 50 ml of plasma with the platelets and label it.

 

 

 

 

 

 

Buffy cot (SAGM) method:

1.    Keep the bags in the buckets of refrigerated centrifuge and balance them accurately. Centrifuge the blood bag at 20-22°C at hard spin (PRP method) for appropriate time.

2.    Extract the plasma in to satellite bag meant for FF P and collect the Buffy cot with approximately 50-60 ml plasma in the small Buffy cot bag.

3.   Transfer the additive SAGM solution to the red cell bag.

4.   Seal and separate the packed red cells and plasma.

5.   Buffy bag along with empty satellite bag is centrifuged at light spin for appropriate time.

6.   Express the Supernatant platelet concentrate to the attached satellite bag.

7.    Leave platelet concentrate undisturbed for 1 hour at 20-22°C, then resuspend the platelet in plasma by gently mixing for 10 minutes.

8.    Store platelet at 20-22°C in platelet incubator with agitator.