1. BLOOD
COMPONENT
THERAPY-II
DR.SAURAV SINGH

3. TERMS
 APHERESIS:
It is a Greek word that means to separate
or to remove. In apheresis,blood is
withdrawn from a donor or patient in
anticoagulant solution and separated into
components. One or more component is
retained and remaining constituent are
returned to the patient.
 PLASMAPHERESIS:
The process of removing the plasma from
red cells is termed plasmapheresis.
Similarly terms are given to removal of
other components like
Platelet(PLATELETPHERESIS), Red

4. PLATELETS
o There are two methods from which
platelets can be obtained:
◦ Differential centrifugation of unit of whole blood
(platelet concentration).
◦ Plateletpheresis
Platelet concentration:
 Platelet concentrate is prepared from centrifugation
of whole blood within 6hrs of donation and is
centrifuged at low spin to produce platelet rich
plasma(PRP)

5. • PRP is transferred to satellite bag and spun at
high speed to get platelet (at bottom) platelet
poor plasma (at top). Platelet poor plasma is
returned to the primary bag leaving behind
50-60 ml of plasma with platelet.
• Platelet is stored at 20-24 degree C, with
agitation which causes exchange of
gases, maintenance of pH and reduces
platelet aggregates and should be used within
5 days.

6. PLATELET CONCENTRATE
RANDOM DONOR
PLATELET(RDP) SINGLE DONOR PLATELETS(SDP)
Prep from whole blood donations Prep by platelet pheresis in cell
Vol-50-60ml separator
Platelets- 5.5x109/bag or more Vol – 150-300ml
Red cells- <1.2x109/bag Platelets – 150-500x 109/bag
White cells- <0.12x109/bag
It may be supplied as single unit
or
Pooled unit of 4-6 donors

7.  Platelets can be stored in bags made up
of Polyvinylchloride(PVC) with Di
(2-ethylhexyle) phthalate(DEHP)
plasticizer up to 72 hrs at room temp.
while certain polyolefin with no plasticizer
maintains the platelet function and pH for
7 days.
 One unit of platelet concentration contains
>45x109 platelet, so transfusion of one
unit raises platelet count by
5000/microlitre

8. Plateletpheresis: single donor is connected to
the blood separator machine in which whole
blood is collected , platelet is separated and
retained while rest of component is returned to
the donor.
This method yields large number of platelet
from single donor(6 units of whole blood) upto
30,000-60,000/microlitre

9. Dosage:
 As a guide, each unit of random donor
platelet should raise platelet in an adult by
5000 to 10000/cumm.
 1 unit /10 kg body weight of RDP is
required
 Bleeding, fever, infection, splenomegaly, al
loimmunization, and intravascular
consumption, each decreases the
expected increment. Response to platelet
infusions should be continually monitored
as essential elements of patient

10. QUALITY CONTROL(QC)
Parameter QC reqd Frequency of
control
Vol 40-70 ml All units
Platelet count >5.5X1010/unit 4 units/month
pH >6.0 -do-
Residual 2X109/unit -do-
leucocytes
RBC Traces-0.5 ml -do-
10

11. o The usual indications are:
• Thrombocytopenia due to decrease in platelet
production like aplastic anaemia ,hematologic
malignancies ,following radiotherapy or
chemotherapy.
• or due to increased platelet destruction like
ITP, DIC,alloimmune thrombocytopenia, drug
induced thrombocytopenia, septicemia.
• Heriditary disorders of platelet dysfunction and
massive blood transfusion.
• Infections like dengue and leptospirosis

12. o Most of the adverse reaction with platelet
transfusion
• is due to presence of leucocytes and plasma
leading to febrile non-haemolytic transfusion
reaction , allergic reaction
• Septicemia due to bacterial contamination
• Alloimmunisation .

13. Leucoreduced platelets
 To minimize the adverse effects of
leucocytes present in blood
components , the use of leukocyte
reduced platelets have been instituted.
 It can be accomplished by two
methods:
 Filtration
 Apheresis component programme.

14. Platelet agitator
Platelet agitators maintain donor platelets in an even suspension throughout the
blood plasma. Platelet agitators are used for storing platelets at a specified
temperature range usually between 20 C-24 C

15. PLASMA COMPONENTS
 FRESH FROZEN PLASMA (FFP):
 Plasma is separated from whole blood by
centrifugation and is transferred to satellite
bag which is rapidly frozen at -80 degree
C and then the temp is brought to -18 to
-30 deg C. This process is done within 6
hrs.
 Storage- FFP can be stored for 1yr at -30
degree C and when required should be
thawed at 37 degree C .

16.  Contents of 1 unit of FFP prepared from
450ml of whole blood.
PLASMA 175 – 250ml
Fibrinogen 200 – 400 mgm
Rich in factor VIII and I ; Labile factor V
 DOSAGE:
About 5-10 units / kg body weight.

17. QC of FFP
Parameter QC Frequency of
control
Volume 200-220 plasma 4 units/month
Stable 200 units of -do-
coagulation each factor
factors
Factor VIII 0.7 units /ml -do-
Fibrinogen 200-400 mg -do-

18.  Indications of FFP:
◦ Deficiency of multiple coagulation factors as in
liver disease, massive transfusion, DIC .
◦ Familial factor V deficiency
◦ Deficiencies of factor II,VII,IX,X
◦ Antithrombin deficiency
◦ Inherited coagulation factor deficiency.

19.  CRYOPRECIPITATE
 Cryoprecipitate is prepared from FFP.FFP
is kept in the refrigerator upside down at -
30 deg C and then at 4-6 deg C ,FFP
melts. This melted FFP moves to another
bag,10-20 ml of FFP left is the cryoppt and
FFP in other bag is cryo poor plasma.
 When required for transfusion, cryoppt is
thawed at 4 deg C in circulating water
bath.

20.  Cryoprecipitate contains factor VIII,von
Willebrand factor, fibrinogen, F XIII and
fibronectin.
 The usual dose of cryoprecipitate in
treating hypofibrinogenemia is an initial
infusion of 10 bags, followed by 10 to 20
bags or as necessary to keep the
fibrinogen level above 100 mg/dl. The
half-life of fibrinogen is about 4days.

21.  Indications:
Used in treatment of Factor VIII deficiency,
von Willebrand disease, F XIII deficiency
and hypofibrinogenaemia.

22. CRYO POOR PLASMA
 This is the supernatant remaining from
the production of cryoprecipitate. It is
relatively deficient in high molecular
weight forms of Von wille brand factor
while retaining normal levels of the
vWF-cleaving metalloprotease.
 Use- Treatment of chronic relapsing
thrombotic thrombocytopenic purpura.

23. Liquid plasma
 Plasma removed from liquid whole
blood up to five days after the expiration
of the whole blood .
 Plasma may be stored in the liquid
state at
I – 60 C.It contains stable clotting
factors, however labile clotting factor
such as factor VIII and factor V are lost.
USE: In deficiency of stable clotting
factor(II,VII,IX,X,XI).

24. Solvent/Detergent treated
plasma
Plasma is treated with the solvent
tri(n-butyl) phosphate (TNPB) and the
detergent Triton X-IOO to inactivate
lipid-enveloped viruses such as
hepatitis B and C and HIV.
 It has no effect on non-enveloped
viruses like hepatitis A and parvovirus
B 19.

25. GRANULOCYTE
CONCENTRATE
Granulocyte concentrate is rarely used
because:
 Most infections are controlled with
antibiotics.
 A granulocyte concentrate prepared
from a single donor has insufficient
granulocyte and contaminated with red
cells.
 Transfusion of granulocyte is associated
with significant risks.
Granulocyte for transfusion can be
obtained single donor unit by differential
centrifugation or by leucapheresis.

26.  Leucapheresis is preferred because it
yields more granulocyte,which can be
further be enhanced by using
corticosteroids.
 Each concentrate contains approximately
10
10 granulocytes which are about one
tenth of the normal adult’s daily production
and that is far fewer than that of an
infected patient. Granulocytes are fragile
and may be stored no longer than 24 h.
The usual concentrate contains about 250
ml of plasma and has a Hct of 15 to 20
percent. ABO compatibility is necessary.

27. Indications:
Patients with severe neutropenia with
documented bacterial or fungal infections.
Patients not responding to antibiotics.
There is evidence that granulocyte
transfusions can benefit a selected group
of patients: those with gram-negative
sepsis or progressive localized infections,
severe granulocytopenia, and temporary
suppression of leukocyte production.

28. BLOOD DERIVATIVES
 HUMAN ALBUMIN
-- Comprised of 96% albumin and 4%
alpha and beta–globulin.
-- Prepared by cold fractionation of pooled
plasma.

29.  INDICATIONS
-- Used as replacement fluid in
therapeutic plasma exchange and
treatment of diuresis resistant edema.
-- In hypovolemic shock, hypotension
associated with hypovolemia in liver
failure or protein losing conditions.

30.  FACTOR VIII CONCENTRATE
-- Prepared by fractionation from large
pools of plasma
-- Heat treated to eradicate any HIV or
hepatitis virus contaminants.
-- Available in freeze-dried forms
 INDICATIONS:
-- Hemophiliacs
-- severe Von Willebrand’s Disease

31. FACTOR IX CONCENTRATE
-- Both plasma derived and recombinant
factor IX concentrates are available.
INDICATIONS:
-- Hemophilia B

32.  PROTHROMBIN COMPLEX
CONCENTRATE
-- Combination of blood clotting factors
II,IX,X and sometimes factor VII as well as
protein C and S.
 INDICATIONS:
-- Inherited deficiency of factor IX,X or II
-- Hemophilia A with inhibitor antibodies
against factor VIII and who are non
responsive to factor VIII concentrate.

33.  IMMUNOGLOBULINS
-- Prepared by cold ethanol fractionation of
pooled plasma.
-- They are of two main types:
 Non- specific immunoglobulins
-- Prepared from pooled plasma of non-
selected donors
-- Composed of antibodies against
infectious agents prevalent in the donor
population

34.  INDICATIONS:
-- Passive prophylaxis against hepatitis A.
-- Congenital or acquired
hypogammaglobulinaemia .
-- Autoimmune thrombocytopenic purpura
to raise platelet count.

35.  Specific immunoglobulins
-- Prepared from donors who have specific
high titer IgG antibodies.
 INDICATIONS:
-- Specific immunoglobulin for passive
prophylaxis against hepatitis B,varicella
zoster,cytomegalovirus,or tetanus.
-- Anti-RhD immunoglobulin used for
prevention of immunization against RhD
antigen in RhD-negative mothers during
pregnancy.

36. RECENT ADVANCES
 Semi automated methods
 Methods for Apheresis
◦ Manual method
◦ Automated methos
 Pharmacological products as
alternative to blood components.

37.  Semi automated
methods:
◦ Preparation of L-R cells
concentrate
◦ Preparation of platelet
from buffy coat
 OPTIPRESS:
automatic extractor
having two plates,one is
stationary and other
expels plasma in empty
satellite bag and red cells
into bag containing
SAGM.

38. OPTIPACKS:
•Contains one 600ml bag
made up of PVC having
63ml CPD phlebotomy
needle is attached. this
bag is connected to two
400ml bags ,one for
collection of plasma and
the other platelets which
can be stored for 5 days .

39. APHRESIS
 Apheresis is collection of anti-coagulated
whole blood from a donor, its separation
into components, retention of desired
component and return of remaining
constituents back to the donor with the
help of automated cell separator
machines.

40. ADVANTAGES OF APHRESIS
 Reduced multiple donor exposure
◦ Reduced risk of alloimmunization
◦ Reduced incidence of transfusion transmitted
diseases
 Full and effective transfusion dose
 Purer product:
◦ leucocyte reduced products
 High quality product
 Fewer donor reaction due to return of fluid

41. Types of cell separators
Intermittent flow cell separator (closed
system)
Continuous flow cell separator
Automated separation techniques by
centrifugation
Cell separation by membrane filtration
 Continuous magnetic cell separator
(immunomagnetic)

42.  Automated separation technique by
centrifugation:
◦ Centrifugal force separates blood into different
component depending upon the specific
gravity.
◦ Blood is drawn from an automatic pump
Anticoagulant is added to the tube and blood
is pumped into rotating bowl chamber in
which layering of components occurs based
on the density. The desired component is
retained and rest returned to donor either by
continuous flow or by intermittent flow.

43.  Separation by Membrane Filtration:
◦ Filtration of plasma through membrane which
allows collection of plasma from a healthy
donor.
◦ Membranes are arranged as hollow fibres
which expels the cellular elements in the flow
of blood.
◦ Most commonly used apheresis devices are:
 Haemonetic corporation: Platelets, plasma,
leucocytes.
 Baxter: Plasma, platelets, red cells, leucocyte
 Gambro: Plasma, platelets, leucocyte and peripheral
blood stem cells.

44.  HAEMONETIC:
It is intermittent centrifuge separator.
The anticoagulated blood is pumped
into rotating bowl .This incoming blood is
separated.The red cells move to the
periphery and plasma to inside of rotating
bowl and the white cells and plasma
between red cells and plasma.
Using optical detectors and fluid surge
elutriation process,the desired component
is retained.

45. HAEMONETICS CENTRIFUGE

46.  GAMBRO(Cobe)
 Continuous flow centrifuge cell separator
where two arm blood is drawn and
returned.
 Here flat membrane is used to separate
the cells of blood from plasma.
 Allows lower WBC and RBC
contamination in platelets.

47.  BAXTER
 Continuous flow technology.
 CS 3000 has two separation containers
firstly for collection of leucocytes reduced
platelets and other for white cells (CS
3000 plus).

48. REFERENCES
 DGHS Manual of blood transfusion
2003
 Essential of clinical Haematology –
Shirish M kawathalkar
 INTERNET

50. THANK YOU