BLOOD TRANSFUSION COMPONENT PREPARATION.ppt

EQUIPMENTS
 Collection Bag :- Double, Triple, Quadruple Bag
 Refrigerated centrifuge
 Plasma Extractor
 Cooling water bath
 Freezer -180
C, -800
C
 Blood Refrigerator 1 - 60
C
 Platelet incubator

Platelet Concentrate Preparation
• Two methods from WB are used;
» Platelet rich plasma (PRP)
» Buffy coat
• Aphaeresis

Platelet Preparation
a)From whole Blood

Platelet Concentrate Preparation
• Initial gentle centrifugation to obtain platetet
rich plasma(PRP)
• 2nd heavy centrifugation to obtain platelet
concentrate
• No special equipment required except a
calibrated centrifuge

Platelet Preparation cont……
• Platelet rich plasma (PRP) is separated from
WB by ‘light-spin’ centrifugation and the
platelets are then concentrated by ‘heavy
spin’ centrifugation, with subsequent removal
of supernatant plasma – platelet poor plasma
(PPP)

Platelet Preparation cont…..
• WB units intended for the preparation of
platelet concentrates must:
» Be maintained between 20-24°C until
platelet concentrate is prepared
» Have the PRP separated from the red cells at
or before 8 hours post collection time

• Place WB in centrifuge making sure that the
temperature is 20°C

• After centrifugation,
place the whole
blood unit into the
plasma extractor and
affix the appropriate
labels to the satellite
bags

PRP –after
1st spin
 Separated PRP from WB with
attached satellite bag

• Place the PRP and attached bag in a plastic
bag and weigh on scale
 Place bags in the centrifuge cup with the
attached bag behind the PRP
 Balance the opposing side(s) of the centrifuge
 Centrifuge the PRP using a ‘heavy spin’ (2nd
spin)

 After centrifugation,
place the spun bag into
the plasma extractor
 Express all but
approximately 55 mL of
plasma (PPP) from the
platelet concentrate
 Close the tubing using a
plastic clamp

RBC-SAGM
PPP
(FFP)
Platelet Concentrate

• Allow platelet
concentrate bag to
‘rest’, label face
down, for 1 hour in a
designated
temperature
controlled area (20-
24°C)

Platelet Preparation cont
b)From buffy coat
• The WB is centrifuged (10 min at 3600 rpm).
• The plasma is pressed into an empty satellite
bag and the buffy coat into the 300 ml bag
which is the 2nd in the row of 4
• The SAGM in the 4th is added to the RBC and
30 ml of plasma is returned to the bag
containing the buffy coat.The RBC and the
plasma are then detached.

Platelet Preparation cont
• The buffy coat is mixed gently with the plasma
and centrifuged for 6 min at 1250 rpm
• The PRP is then slowly transferred to the bag
which originally contained the SAGM.
• The resuspended buffy coats (which should
ideally have been previously pooled) will
provide an adult platelet dose of 3 x 1011
platelets in ~ 300 ml of plasma

Platelet storage
• Place the platelet
concentrate onto the
appropriate platelet
rotator/reciprocator
for storage at 20-
24°C with
continuous agitation
Platelet agitator

Platelet Preparation
c) Platelet Pheresis
 Trima – device used to
collect Aphaeresis
Platelets
 Adult dose of platelets
from single donor

Preparation of Cryoprecipitate
• Cryoprecipitate as also called Antihemophilic
factor (AHF)
• AHF (Coagulation Factor VIII) can be
concentrated from freshly collected plasma by
cryoprecipitation
• Cryoprecipitation is accomplished by slow
thawing frozen plasma (FFP) at 1 to 6 degrees
C

Preparation of Cryoprecipitate cont....
 Fresh WB collected in a unit with at least 2
integrally attached satellite bag(s)
 Refrigerated centrifuge and plasma extractor
 Plastic clamps, heat sealer, and tube stripper
 Refrigerator (1-6°C)
 Freezing apparatus (-18°C or colder)

Preparation of Cryoprecipitate cont...
 Collect WB in a triple or quadruple blood unit
 Make sure the blood is kept at 4°C during
transport and before processing steps begin

 Centrifuge the WB shortly after collection at
4°C
 Place the plasma bag in a freezing apparatus
(-180°C)
 The 1st spin will separate the red cells and
plasma
 The 2nd spin (following the plasma thawing)
will separate the cryoprecipitate from the
cryoprecipitate-poor plasma

 Place the centrifuged WB in a
plasma expressor, open the seal
and express the plasma into the
attached satellite bag until the
plasma red cell interface reaches
the primary port (RBC-SAGM). If
RBC-CPDA unit, leave 3 cm of
plasma.
 Clamp the tubing next to the
plasma bag with a plastic clip.
 Before separating the plasma
from the primary bag, ensure
that satellite bag is labeled with
correct donor number

 Heat seal and detach
plasma
 Affix the appropriate
cryoprecipitated
label on the front of
the plasma bag

 If appropriate, add
SAGM to the RBC and
mix well. Heat seal and
detach SAGM bag
 Save the empty SAGM
bag and do not detach
 Complete specific RBC
preparation in
accordance with SOP

 Promptly place plasma
in a freezing device so
that freezing is started
within the time frame
required
 The plasma and the
satellite bag should be
placed on a flat surface
in the freezer SAGM or other satellite bag Plasma

 After at least 24 hours,
remove the FFP from
the freezer and place on
a shelf in the
appropriate refrigerator
 Allow the FFP to thaw
slowly at 4°C 12-15
hours
 When the plasma has a
slushy consistency,
remove from the
refrigerator and towel
dry the bag and tubing

• Centrifuge the unit in an upright position in a
calibrated centrifuge at 4°C for the time and
rpm noted on the centrifuge
• Two plasma units can be put into one
centrifuge cup

 After centrifugation,
place the plasma in
the expressor
 Allow the
supernatant plasma
to flow slowly into
the satellite bag
 Process no more
than 8 units at a
time

 The ice crystals at
the top of the bag
serve as a filter

 Do not allow any cryoprecipitate to express
with the plasma
 The cryoprecipitate paste will adhere to the
bottom of the bag or to the ice

 Temporarily clamp the tubing with a plastic
clip when about 90% of the cryoprecipitate-
reduced plasma has been removed
 Leave approximately 10-15 mL of supernatant
plasma in the cryoprecipitate bag
 Heat seal the tubing between the bags and
separate
 Attach the proper component labels to the
blood products

 Refreeze the prepared cryoprecipitate
products immediately
 The entire process (removing the thawing
plasma from refrigerator → to refreezing the
prepared cryoprecipitate) should not take
longer than one hour

• Frozen component is thawed in a protective
plastic over wrap in a water bath at 30-37°C
up to 15 minutes
• Do no refreeze after thawing
• Thawed Cryoprecipitated AHF should be kept
at room temperature and transfused within 6
hours if it is a closed single unit or within 4
hours if it is an open system or units have
been pooled

• For pooling, the precipitate in each
concentrate should be mixed well with 10-15
ml of diluent to ensure complete removal of
all material from the container.
• The preferred diluent is0.9% Sodium Chloride,
Injection. Several units can be pooled and the
volume of the pool is indicated on the label

Frozen RBC
• Blood is frozen to preserve: rare types, for
autologous transfusion, stock piling blood for military
mobilization and/or civilian natural disasters.
• Blood is drawn into an anticoagulant preservative.
– Plasma is removed and glycerol is added.
– After equilibration unit is centrifuged to remove
excess glycerol and frozen.
• Storage temperature
– high glycerol -65 C.
– low glycerol -120 C, liquid nitrogen
• Expiry
– If frozen, 10 years.
– After deglycerolization, 24 hours.

Frozen RBCs; Deglycerolized RBCs
• Thaw unit at 37C, thawed RBCs will have high
concentration of glycerol.
• A solution of glycerol of lesser concentration of
the original glycerol is added.
• This causes glycerol to come out of the red
blood cells slowly to prevent hemolysis of the
RBCs.
• After a period of equilibration the unit is spun,
the solution is removed and a solution with a
lower glycerol concentration is added.
• This procedure is repeated until all glycerol is
removed, more steps are required for the high
glycerol stored units.
• The unit is then washed.

Rejuvenated Red Blood Cells
• A special solution is added to expired RBCs up
to 3 days after expiration to restore 2,3-DPG
and ATP levels to prestorage values.
• Rejuvenated RBCs regain normal
characteristics of oxygen transport and
delivery and improved post transfusion
survival.
• Expiration is 24 hours or, if frozen, 10 years

Washed Red Blood Cells
• Washing removes plasma proteins, platelets, WBCs
and micro aggregates which may cause febrile or
urticarial reactions.
• Patient requiring this product is the IgA deficient
patient with anti-IgA antibodies.
• Prepared by using a machine which washes the cells 3
times with saline to remove and WBCs.
• Washed RBCs removes of 85% of WBCs.
• To be used within 24 hours
No longer considered effective method for
leukoreduction.

RBCs Leukocyte Reduced
• Leukocytes can induce adverse affects during transfusion,
primarily febrile, non-hemolytic reactions.
• Reactions to cytokines produced by leukocytes in transfused
units.
• Other explanations to reactions include: immunization of
recipient to transfused HLA or granulocyte antigens, micro
aggregates and fragmentation of granulocytes.
• Historically, indicated only for patients who had 2 or more febrile
transfusion reactions, now a commonly ordered, popular
component.
• “CMV” safe blood, since CMV lives in WBCs.
• Most blood centers now leukoreduce blood immediately after
collection.
• Bed side filters are available to leukoreduce products during
transfusion

Irradiated RBCs
– Prevents T-cell proliferation that may cause
transfusion-associated graft versus host disease
(GVHD)
– GVHD is fatal in 90% of those affected
– Used for:
• Donor units from a blood relative
• HLA-matched donor unit
• Intrauterine transfusion
• Immunodeficiency
• Premature newborns
• Chemotherapy and irradiation
• Patients who received marrow or stem cells

Clinical Use of Blood Components

WHOLE BLOOD (WB)
Most blood transfusions are not WB
Indications
 Symptomatic anemia with large volume loss –
e.g., massive transfusion
 Exchange transfusion
Benefits
 Increases oxygen and blood volume
 WB must be ABO identical

RED BLOOD CELLS
(RBC-CPDA/SAGM)
Benefits
 Increases oxygen-carrying capacity
 Must be ABO-compatible
 RBC-CPDA sometimes preferred

RED BLOOD CELLS
(RBC-CPDA preferred
 Higher % of red cells to total volume of
product infused – pediatric transfusion
 Concern regarding potential side effects of the
adenine, dextrose, and mannitol in AS
solutions
 Decision may also be dependent on
availability of product

RBC
• Conditions include:
– Symptomatic anemia;
– exchange transfusion
– Oncology patients (chemo/radiation)
– Trauma victims
– Cardiac, orthopedic, and other surgery
– End-stage renal disease
– Premature infants
– Sickle cell disease ( Hgb A)

Fresh Frozen Plasma (FFP)
FFP serves as a source of plasma proteins for
patients who are deficient in or have defective
plasma proteins
Indications
 Management of bleeding or preoperative
patients who require replacement of multiple
coagulation factors (e.g., liver disease)
 Massively transfused patients with clinically
significant coagulation deficiencies
 Patients on warfarin who are bleeding or need
to undergo surgery

PLATELET CONCENTRATE
Indications
 Bleeding due to thrombocytopenia
 Bleeding due to platelet function abnormality
 Prevention of bleeding in marrow hypoplasia

Platelet Concentrate
Benefits
 provide adequate numbers of normally
functioning platelets for the prevention and
cessation of bleeding
 Platelets not indicated in patient’s with TTP or
ITP

CRYOPRECIPITATED AHF
Indications
 Hemophilia A (factor VIII deficiency)
 von Willebrand’s disease
 Hypofibrinogenemia
 Factor XIII deficiency

CRYOPRECIPITATED AHF
Benefits
 Serves as a source of Factor VIII, fibrinogen,
vWF, and Factor XIII
 If virus-inactivated Factor VIII concentrates or
recombinant factor concentrates are available,
cryoprecipitate should not be used

PLASMA CRYOPRECIPITATE REDUCED
Indications
 Transfusion or plasma exchange in patients
with TTP that are refractory to FFP
 Provide clotting factors except Factor I
(fibrinogen), VIII, XIII, and vWF

PLASMA CRYOPRECIPITATE REDUCED
Benefits
 Source for deficient plasma proteins – Factors
II, V, VII, IX, X, XI and albumin
 This component should not be used as a
substitute for FFP

BLOOD TRANSFUSION COMPONENT PREPARATION.ppt

Recommended

Recommended

More Related Content

Similar to BLOOD TRANSFUSION COMPONENT PREPARATION.ppt

Similar to BLOOD TRANSFUSION COMPONENT PREPARATION.ppt (20)

Recently uploaded

Recently uploaded (20)

BLOOD TRANSFUSION COMPONENT PREPARATION.ppt