Blood Components

Dr. JEEWAKA GALHENAGE

MBBS, DTM, MD (Transfusion Med)
CONSULTANT TRANSFUSION PHYSICIAN
• Introduction
• Preparation
• Storage Quality Control
• Indications
• Effective blood transfusion therapy depends upon the availability of
different blood components.
• blood donor donates whole blood, from which different components
are prepared.
• Advantages of component preparation
1. Optimal survival of each component
2. Transfusion of the only specific blood component required by the
patient.
3. A single blood donation can provide transfusion therapy to
multiple patients
4. Minimize unwanted effects of transfusion
 Blood Products

Whole blood
• One unit of donor blood collected in a 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 unit or by apheresis

Blood Derivatives ( Plasma derivatives)

A product obtained from multiple donor units of plasma by

fractionation.
 Red Cell Concentrate ( Packed Cells)

Leucodepleted/ Leucoreduced red cells

Whole
Blood Platelet Concentrate ( Random Donor Platelets
Derived
Lecoreduced platelet Concentration

Cellular Granulocyte Concentrate

components
Erythrocytapheresis
Apheresis
Derived Platelet Apheresis
Granulocytes Apheresis
Peripheral blood stem cells
 Fresh Frozen Plasma (FFP)

Single donor plasma

Plasma components
Cryoprecipitate

Cryo-poor plasma
 Albumin

Factor VIII concentrate

Factor IX concentrate

Plasma derivatives Prothrombin complex concentrate

IV Immunoglobulins

Fibrinogen

Other Iv Ig / Coag. Fac /

Preparation of Blood Components
Equipment
1. Refrigerated blood bank centrifuge
2. Blood bank refrigerator
3. Freezer –20º C or below
4. Platelet agitator
5. Weighing scale for balancing centrifuge cups
6. Tube sealer
7. Plasma Defroster or Water Bath
8. Plasma expresser (manual)
9. Component Separator
10. Stripper/ Cutter
Consumable Items

1. Sealing Clips
2. Double, Triple and quadruple blood bags
3. Appropriate sticker type labels and instructions for each component
4. Gloves
Methods of Preparation

• There are various methods to separate

• Yield and quality of component depends upon the method applied.

Various methods used are; -

(a) Gravity separation - crude method
(b) Low and high speed refrigerated centrifugation
• Due to different specific gravity of cellular components, they can be
separated by centrifuging at different centrifugal force (in g) for different
time.
(c) Apheresis by cell separator.
Precautions to be observed in preparing components

In collection of blood:
• Proper selection of donor
• Clean and aseptic venepuncture site with minimum tissue trauma
• Uninterrupted and continuous flow of blood
• Collection of correct amount of blood proportionate to anticoagulant
(450ml+/- 10%)
• Proper mixing of blood during collection to mix with anticoagulants
• Platelets should be separated within 6-8 hours from the time of
collection of blood.
In centrifugation:

• Opposing cups with blood bag and satellite bags must be equal in
weight (Proper balancing)
• Correct speed of centrifugation, time and temperature must be
maintained.
• Follow aseptic techniques & adhere to SOP
Blood Bags

• Made with high molecular weight PVC to ensure better tensile

strength and weld strength
• Designed for the collection, processing and storage of whole blood
and blood components.
• It acts as a closed system reducing the chances of contamination
• Types of bags – Single, Double, Triple, Quadruple, Transfer Bags
Single Blood Bags

• For Whole Blood Collection

• Bag contain CPDA as anticoagulant
Double Blood Bags

• For Whole Blood Collection

• Separation of two different blood components (Red Cells & Plasma)
obtained through the process of centrifugation
Triple Blood Bags

• Triple Blood Bags with SAGM for Whole Blood collection

• Separation of Three different Blood Components ( Red Cells,
Plasma & platelets)
• The primary bag contain CPD and one satellite bag contains
SAGM
Quadruple Blood Bags
(Top & Top / Top & Bottom)

• For whole Blood Collection and separation of four blood components -

Red Cells / platelets / Plasma/ Buffy Coat (white cells)
• Primary bag contain CPD and one satellite bag contain SAGM solution
for red cell preservation.
Anticoagulation and Preservation

Common Anticoagulant Solutions

• CPD (Citrate Phosphate Dextrose)

• CPDA (CPDA-1 and CPDA-2) (Citrate Phosphate Dextrose
Adenine)
• ACD (Acid Citrate Dextrose)

All above citrate based anticoagulants prevent coagulation by chelation

of calcium
ACD Solution

• Preserves ATP level

• Helps maintain red cells shape.
• Prevents haemolysis
• Maintains pH
• Shelf life of whole blood / red cells in ACD = 21 days
CPD Solution

• Decreases acidosis
• Improves ATP synthesis
• Shelf-life of whole blood in CPD = 28 days
Citrate Phosphate Dextrose Adenine (CPDA):
Adenine
• Helps maintain high ATP levels
• Blood collected in CDPA
• safe
• well tolerated
• 2,3 DPG levels can be maintained for 12-14days
• shelf-life = 35 days
Additive Solutions
• These solutions contain saline, adenine and glucose and are added to
the red cells after separating them from plasma. ( Preserving Solution)

The advantages of using additive solution

1. Better storage condition for red cell preparation and lowering of
viscosity for ease of transfusion.
2. Increased yield of plasma for plasma fractionation.
3. Removal of unwanted buffy coat
4. Additional 7 day storage time for red cell preparation
5. Avoid unnecessary transfusion of plasma
Common Additive Solutions

• SAGM- This additive solution has Sodium Chloride, Adenine,

Dextrose & Mannitol to prevent lysis of red cells. The mean 24 hr.
post transfusion survival is better with the additive.

• .ADSOL- This additive solution is similar to CPD-SAGM but has

greater quantities of glucose, adenine and Mannitol. It has better red
cell preservation and 24 hr post-transfusion survival. The shelf life of
Red Cells with this solution can be increased to 42 days.
Preparation of Blood Components by Centrifugation
 Double Bag
Heavy spin at 4⁰C within 8 hours

Packed Red Cells Fresh Plasma

Freeze -50 ⁰C
Stored at 2 -6⁰C immediately

Stored at ≤-30⁰C
 Triple Bag
Light spin at 22⁰C within 8 hours

Packed Red Cells Platelet Rich Plasma

Heavy spin at 22⁰

Platelet Fresh Plasma

concentrate

Freeze
Stored at 22⁰C Stored at ≤-30⁰C
 Quadruple Bag

RCC BC removed Buffy Coat FFP

in AS

Platelet concentrate

Buffy coat
Whole Blood

• Blood taken from a suitable donor using a sterile , pyrogen free

anticoagulant & container
• Source material for component preparation ( Usually not stored in
blood banks as all the units used for component separation)
• Volume – 450 ml+/- 10% Hb – 45 g / unit Stored at 2 C – 6 C
• Storage period depends on anticoagulants used ( CPDA-1, 35d )
• No labile clotting factors / No functional Platelets
Red Cell Concentrate – RCC ( Plasma Suspended)

• A unit of concentrated red cell prepared from whole blood by

centrifugation & removal of part of the supernatant plasma

• Stored at 2 C – 6 C and storage period depends on the

anticoagulants used ( Usually in CPDA-1 for 35 d)
 Red Cell Concentrate
(Buffy Coat removed in Additive Solution)

• A red cell component prepared by the removal of a major part of the

plasma and the buffy coat layer from whole blood with subsequent
addition of an appropriate nutrient solution.

• Stored at 2 C – 6 C

• Storage period depends on the anticoagulants/ Additive system used

( Usually in CPDA-1 with Adsol solution for 42 d)
 Quality Monitoring
Red Cell Concentrate (Buffy Coat removed in Additive
Solution)

Parameter Specification

Volume 230 – 330 ml

% Hct 50 -70 %
Hb > 43 g/Unit
pH > 6.4
Haemolysis at EOS < 0.8%
WBC Count 1 x10 9 /Unit
 Leucoreduced Red Cells (LRRC)
• < 1x 109 WBCs

• Use of BC method for component

production using top-bottom bags

• Besides red cells even platelets are

leucoreduced.

• Used in FNHTR / Mild allergic

conditions
 Leucocyte Depletion (LD)
LD blood components = those containing <5x106 WBCs
per unit
Prepared by leucocyte depletion filters
Components required to be leucodepleted: WB, PRC,
RDP and SDP
Filtration can be done :

• Pre-storage - during component processing

• Before issue
• Bedside
 LD Blood Components

INDICATIONS

• To reduce risk of alloimmunisation in multitransfused

patients

• To reduce risk of transmission of CMV, EBV and

HTLV-I

• If FNHTR to red cell transfusion

• May help in prevention of immunosuppression, which

leads to postoperative infection and tumour recurrence
(evidence inconclusive)
Red Cells - Washed

• A red cell component prepared by the centrifugation of the primary

component and removal of plasma or additive solution (and if
applicable buffy coat layer ) then washing by sequential addition and
removal of an additive ( usually saline) solution.

• Most of the plasma, leucocyte & platelets are removed.

• Must be kept at controlled temp between 2 C – 6 C
• Storage time 6 h ( must never exceed 24h)
 Quality Monitoring
Red Cell Concentrate (Washed)

Parameter Specification

Volume 230 – 330 ml

% Hct 65 – 75%
Hb > 40 g/Unit
pH > 6.4
WBC Count < 1 X10 9/unit
INDICATIONS for Washed Cells

• Prevention of FNHTR (Febrile Non-Hemolytic Transfusion

Reactions)
• Prevention of allergic and anaphylactic reactions
• For Ig A deficient patients with IgA antibodies
Red Cells – Cryopreserved ( Frozen Red Cells)
• Red cells are frozen, preferably within 7 days of collection, using
cryoprotectant, and stored at – 60 C to – 80 C or below depending on
the method of cryopreservation

• In general, two methods;

1. High glycerol- Storage temp -60 C to -80 C
2. Low glycerol – Storage temp -140 C to -150

Both methods require a washing / deglycerolisation procedure

Storage can be extended up to 10 yrs
• Rare blood group / phenotype units are frozen to use in needed pts.
Granulocytes (Buffy Coat)

Prepared by
9
1.From WB – Buffy Coat Method >1x10
• Therapeutic dose - 10 packs (1.0 x 1010 daily/bd)
2.Aphaeresis technique- Granulocyte concentrate
≥ 1.0 x 1010 ( therapeutic dose)
• Needs irradiation
• Storage at room temperature (22oC)
• Shelf life 24hrs
• Cross-match to be done
 CLINICAL INDICATIONS FOR GRANULOCYTE
TRANSFUSIONS

Clinical effectiveness is controversial

Pt who fulfil all of the following criteria:
1. Severe neutropenia, defined as ANC <0.5 x109/L due to congenital
or acquired bone marrow failure syndromes
2. Proven or highly probable fungal or bacterial infection that is
unresponsive to appropriate antimicrobial therapy
3. In whom neutrophil recovery is expected (ANC>0.5x109 /l)
FFP ( Fresh Frozen Plasma)

• FFP is separated from whole blood collected using a blood bag with
integral transfer packs preferably with in 6 – 8 h of collection
• Storage temperature: – 30°C Shelf Life- 1 Year
• Indication- 1. patients with bleeding due to multiple clotting factor
deficiencies such as DIC
2. inherited clotting factor deficiencies (e.g. Factor V
deficiency) where a clotting factor concentrate is
not yet available.
 Quality Monitoring
FFP ( fresh Frozen Plasma)

Parameter Specification
Volume 150 -250 ml

F VIII 0.7 / ml

Fibrinogen > 300 mg /Unit

Preparation of Platelet Concentrate (PC)
Platelet concentrates can be prepared from:
a) Random donor platelet (prepared from whole blood)
b) Single donor platelet prepared by apheresis using various cell
separators
Random Donor Platelet
It is prepared from whole blood kept at room temperature (20-24°C) and
within 6 to 8 hours of collection. As mentioned above for preparation of
platelets from WB, two primary methods are available:
1. Preparation from buffy coat
2. Preparation from PRP.
Platelets prepared from any method:

• Storage temperature -22°C +/- 2°C

• Shelf Life - 5 days
• Must be kept under constant agitation till
used. ( In a platelet agitator)
Platelet Concentrate by
Apheresis (PC –AP)

• The platelets are selectively separated from the whole blood and
retained in the collection bag manufactured specially for platelet
storage.
• The platelet yield is related to donor’s platelet count, the amount of
blood processed and the volume of the product collected.
• Useful specially when providing rare group PLT & supplying HLH
matched PLT
 Quality Monitoring
Platelet Concentrate by apheresis (PC –AP)

Parameter Specification
Volume 150 - 300 ml
Swirling Present
Platelet Count > 200 X 10 9 / Unit
RBC Count < 1 X 10 9/Unit
WBC Count < 1 X 10 9/Unit
pH > 6.4
 Quality Monitoring
Platelet Concentrate (PC –PRPD)

Parameter Specification
Volume 45 – 75 ml
Swirling Present
Platelet Count > 55 X 10 9 / Unit
RBC Count < 1 X 10 9/Unit
WBC Count <0.2 X 10 9/Unit>
pH > 6.4
 Quality Monitoring
Platelet Concentrate (PC –BCD)
Parameter Specification
Volume 45 – 75 ml
Swirling Present
Platelet Count > 55 X 10 9 / Unit
RBC Count < 1 X 10 9/Unit
WBC Count < 0.05 X 10 9/Unit
pH > 6.4
 Cryoprecipitate

• A component containing the cryoglobulin fraction of plasma obtained

by further processing of FFP
• Contain FVIII, FXIII , von Willebrand factor, Fibrinogen &
Fibronectin.
• Prepared by thawing of FFP either overnight at 2C – 6C or by the
rapid thaw- siphon thaw technique. After thawing component is re-
centrifuged using hard spin at same temp. The supernatant cryo –
poor plasma is removed and precipitated cryo is rapidly frozen.
• Shelf life is 1 year at -30 C
-30 C
 Reconstituting cryoprecipitate
(Thawing and issue of Cryoprecipitate)

• Place in a 37°C water bath until the cryoprecipitate has dissolved.

• After thawing, re-suspend injecting 10 ml od N.saline in to the bag
(under laminar flow)
• Once re-suspended pool the cryoprecipitate from all thawed bags into
one bag (under laminar flow)
• Once thawed, cryoprecipitate should be kept at 2-6°C and
administered within 4 hours. It should not be frozen.
 Quality Monitoring
Cryoprecipitate ( Singer Donor Pack - Plasma Free)

Parameter Specification
Volume < 5 ml
F VIII > 70 IU / Pack
Fibrinogen > 140 mg /Pack
• Indications

• As Source of fibrinogen(Massive Blood Transfusion

etc)

• Factor VIII deficiency (Hemophilia A)

• Von Willebrand’s Disease

 Cryosupernatant Plasma (CSP)

• Remaining plasma after preparation of cryoprecipitate

• Average volume approximately 200 mL.
• All plasma proteins, FIX, other stable coagulation
factors
• Indications:
• TTP and HUS
• Hypoproteinemic oedema – NS, Burns
,Hypoproteinemia
• Hemophilia B
• Plasma exchange ( TTP)
58
 Irradiation

• The major technology for preventing TA-GvHD is irradiation of blood

components to inactivate residual lymphocytes.

• Blood components that needs irradiation -

Red cells, platelets and granulocytes

• Irradiation not needed for FFP/Cryo/CSP or plasma

TA-GvHD

• Rare but serious complication of BT

• Engraftment & proliferation of donor T lymphocytes, and interaction

with recipient cells expressing HLA Ags causing cellular damage.

• i)Immunocompetent T Lympocytes. ii) Histocompatibility difference

between graft & recipient iii) Usually immunocompromized pt
• Cellular damage to skin ,liver, GIT, Spleen , Bone Marrow
Blood Irradiator
Plasma Derivatives ( Fractionated Blood Products)
• These are licensed medicinal products manufactured from human
plasma donations.
• Products are manufactured from large donor pools.
• Product is selectively precipitated using variations in the
concentration of ethanol, salt, temperature and pH. (the Cohn
Process)
• Multiple pathogen inactivation steps are followed to eradicate
transfusion-transmitted viruses.
Human Albumin Solutions
Clotting Factor Concentrates
Immunoglobulin solutions
Human Albumin Solutions

• Manufactured by cold ethanol fractionation from a large pool of

plasma
• Isotonic solutions (4.5 and 5.0% ) - Often used to replace sub acute
plasma volume loss caused by burns, pancreatitis or trauma, and as a
replacement fluid in plasma exchange.
• Concentrated solutions (20% ) - Used to initiating diuresis in
hypoalbuminaemic patients with liver cirrhosis or nephrotic syndrome,
removal of large volumes of ascites in patients with portal
hypertension exchange transfusion in the newborn
Clotting factor concentrates

• Single-factor concentrates are available for the treatment of most

inherited coagulation deficiencies except Factor V and Factor II
(prothrombin).
• Most patients in the UK with severe haemophilia A are now treated
with recombinant Factor VIIIc ( No viral risk)
• Prothrombin complex concentrate (PCC) contains Factors II, VII, IX
and X. It has replaced FFP in UK as the recommended treatment for
rapid reversal of warfarin overdose, with elevated INR)and severe
bleeding, in view of its superior efficacy,
Immunoglobulin solutions

Normal immunoglobulin:
• Contains antibodies to viruses that are common in the population.
• IM normal immunoglobulin may be used to protect susceptible
contacts against hepatitis A, measles or rubella.
• High-dose IV immunoglobulin is used as replacement therapy in
patients with severe immunoglobulin deficiency & autoimmune
diseases ( ITP)
Specific immunoglobulin:
• Made from selected donors with high antibody levels to the target of
treatment.(tetanus, hepatitis B , rabies & anti –D )
Thank You