BLOOD, CELLULAR COMPONENTS AND

PLASMA PRODUCTS PREPARATION AND

THEIR USES.

DR. RANSFORD KYEREMEH

Aim
To show students how to collect and process donor
blood for use in blood transfusion therapy.

Objectives
 At the end of the lecture students are expected to
know the various donor blood components.

They are expected to be conversant with how to

process, preserve and store these blood units.

They should have a fair knowledge of the uses of

the blood components.
LECTURE OUTLINE
Whole blood.
Concentrated red blood cells (CRC).
Fresh frozen plasma (FFP).
Random–donor platelets concentrates.
Cryoprecipitate.
Factor VIII concentrate.
Factor IX concentrate.
Human leukocyte antigens(HLA)-matched platelets.
Granulocyte concentrates.
Albumin.
 WHOLE BLOOD
One unit of whole blood contains approx. 450 ml of
blood plus 63 mls of anticoagulant (CPD,CPDA-1
etc.).

Though no components have been physically

removed, levels of labile coagulation factors V and
VIII are significantly reduced during storage.

Platelets and granulocytes lose viability in whole

blood stored longer than 24 hrs.
 APPROPRIATE USES OF WHOLE BLOOD
In an adult, one unit of whole blood will increase the Hb by
1g/dl.

Whole blood is used to;

1. improve O2-carrying capacity and for vol.expansion.

2. for actively bleeding patients (loss of vol.>35%).

3. for exchange transfusion in sickle cell disease patients.

DONOR BLOOD UNIT(WHOLE
BLOOD)
 CRC

This normally has a volume of 250-350ml and haematocrit

of 50%-80% depending on the type of anticoagulant used.

CRC does not contain functional platelets or granulocytes or

significant levels of coagulation factors, though the O 2-
carrying capacity may be intact.

CRC is given to patients with chronic symptomatic anaemia.

 CRC cont.
Patients who are actively bleeding (not exceeding
25%) e.g. trauma, surgery, spontaneous hemorrhage.

CRC is not indicated when the total volume loss may

be safely restored by the use of crystalloid (electrolyte)
solutions alone.

A general guideline is to use such solutions up to 25%

of the total blood volume.
 PREPARATION OF CRC
1. Weigh and balance each unit in a centrifuge bucket.

2. Centrifuge whole blood at a high speed (3000rpm) for 20

min. at 2-8oC (refrigerated centrifuge)

3. Using an expressor, express approx. 150 to 250ml of

plasma to yield CRC of HCT. 70-80%.

4. Seal the tube with an electric heat sealer at three different

places and store the plasma at -40oC as FFP.
FRESH FROZEN PLASMA (FFP)

This is prepared from whole blood by separating and

freezing the plasma (200-250ml) within six hours of
phlebotomy.

Contains albumin,globulins,coagulation factors and

other proteins along with water and electrolytes.

Plasma maintained in the frozen state will contain

both labile coagulation factors (V and VIII) and stable
factors (I,II,VII,IX,X,XI,XII,XIII and others).
FRESH FROZEN PLASMA
 FFP USES

Multiple coagulation deficiencies; PT and APTT as in patients

with bleeding Liver dx. and in DIC.

FFP may be indicated in massive transfusion ( when vol. of blood

equal or greater than that of the patient) is given followed by
resuscitation fluids and blood components deficient in
coagulation factors.

This may result in dilutional coagulopathy.

To reverse the effect of warfarin in order to carry out an urgent

procedure since oral vit. K (which is usually used ) takes 24
hours.
 FFP USES cont.

FFP is used to correct a specific coagulation factor

deficiency , such as antithrombin III ,factor
II,V,VII,IX,X,XI or protein C and S.

FFP is used to replace plasma in patients with TTP-

simple infusion of FFP may be of some benefit if
plasma exchange is not possible.
RANDOM–DONOR PLATELETS CONCENTRATES.

Platelets are separated from single units of whole blood

and suspended in small amount of plasma.

Each bag should contain at least 5.5 x10 10 platelets in

50-70ml plasma.
 PLATELET CONCENTRATE PREPARATION

Platelet concentrate can be produced during the normal

CRC preparation or by apheresis.

1. Maintain whole blood (taken for not >6hrs.) at 20-24 oC

before and during platelets preparation.

2. Set the centrifuge temp. to 22oC and spin shortly at a

light spin (depending on the type of centrifuge, about
3200rpm for 2-3min).
 PLATELET CONCENTRATE PREPARATION cont.

4. This process should be done in a closed, multibag

system.

5. Express off the platelets-rich plasma into one of the

satellite bags.

6. Enough plasma must remain on the RBCs to maintain

a HCT. of 70-80%.

7. Seal the tubing between the RBCs and the plasma.

PLATELET CONCENTRATE PREPARATION cont.

6. Disconnect the RBCs and store at 4oC.

7. Recentrifuge the platelet-rich plasma at 22 oC at a higher

speed (approx. 3600rpm for 5min).

8. This will separate the platelets from the plasma.

 PLATELET CONCENTRATE PREPARATION cont.

11. Express the majority of the plasma into the second

satellite bag, leaving approx. 50-70ml on the platelets.

12. The vol. is important to maintain the pH above 6.2.

13. Platelets are resuspended in plasma, allowed to lie on the

bench undisturbed for a while, and kept at 20- 24 oC on a
shaker (shelf life is 5days).
 PLATELET USES
Prophylactically- for non-bleeding patients with <15-20 x
109 platelets or lower. E.g. patients with bone marrow
hypoplasia as a result of chemotherapy, tumor invasion etc.

For patients with ecchymosis, petechaiae,epistaxis,CNS or

GIT bleeding.
 PLATELET USES cont.

Prophylactically before surgery or invasive procedures

such as spinal tap or insertion of central line in patients
with count s below 50-60 x109.

For patients with documented abnormal platelets function

before invasive procedures or to treat bleeding.
 CRYOPRECIPITATE
This is prepared by slowly thawing one unit of FFP at 4 oC(1-
6oc).

The cold insoluble precipitate, resuspended in 10-15 mls of

plasma contains approx:

i. 80-120 units of factor VIII:C (the coagulant

property of factor VIII).

ii. 200 mg of fibrinogen.

iii. 40-70% of von Willebrand factor (VII:vWF).

 CRYOPRECIPITATE cont.

Shelf-life is 12months when frozen and must be transfused

within 6hrs. after thawing and 4 hrs. after pooling( if not
meant to be stored)
 CRYOPRECIPITATE PREPARATION
The whole blood must be cooled since platelets are not
normally prepared from the same unit.

1. Venipuncture must be non-traumatic.

2. At least 200ml plasma should be used to ensure a yield of at

about 80AHF( antihaemophilic factor).

3. The plasma must be frozen within 8hrs of collection and 1hr

from the time freezing starts.
4. The plasma is then allowed to thaw slowly in a refrigerator at
1-6o C.
 CRYOPRECIPITATE PREPARATION cont.
5. Centrifuge the plasma at 4oC at 2000 rpm for 10 minutes
and express the supernatant plasma into the attached
satellite bag.

6.The small white mass in the original plasma bag (10-

15mls) is the cryo and must be separated and frozen
immediately ( to avoid decrease in activity of factor VIII).
USES OF CRYOPRECIPITATE
Treatment of vW dx.

Hypo-fibrinogenemia.

Bleeding or prophylaxis in hemophilia A dx.

(factor VIII:C deficiency)

Factor XIII dx.

Bleeding related to renal failure.

 FACTOR VIII CONCENTRATE

Factor VIII (antihemophilic factor; factor VIII:C)

concentrate is prepared by the fractionation of pooled
plasma.

The quantity of factor VIII is expressed in international

units (IU): 1ml of normal plasma contains 1 IU of factor
VIII activity.
 USES OF FACTOR VIII CONCENTRATE

To control or prevent hemorrhage in patients with hemophilia

A who have moderate to severe congenital factor VIII
deficiency.
 FACTOR IX CONCENTRATE(PROTHROMBIN
COMPLEX )

Factor IX concentrate is prepared by fractionation of plasma

and contains coagulation factor IX along with traces of
factors II,(prothrombin),VII, and X.

Used in treatment of factor IX deficiency (Hemophilia B or

Christmas dx).

Patients with congenital deficiencies of factors VII and X.

 BLOOD BANK
ANTICOAGULANTS

Storage
ACD-Acid-Citrate-Dextrose 21days

CPD-Citrate-Phosphate-Dextrose 28 days

CPDA-Citrate-Phosphate-Dextrose-Adenine 35 days

SAGM-Saline-Adenine-Glucose-Mannitol 42 days
LECTURE SUMMARY