Dr. Kabita Chatterjee (MD)
Faculty-In- Charge
Blood Bank (Main Hospital)
A.I.I.M.S., New Delhi – 110029.
2
Countries where the Buffy Coat production method is
predominantly used for the preparation of platelets
derived from who...
• A layer of mixed white cells and
platelets created by high speed
centrifugation of whole blood
• It is neutral or buff i...
PC FFP
PRP method
PC
Buffy coat method
PR
P
Plasma
RBC
PRP
RBC
Plasm
a
5
Platelets prepared through
buffy method
Activated Platelet
prepared through PRP
method
PLATELET – Buffy coat V/s PRP
Plate...
 In recent years, platelet rich buffy coat (BC) has
become an alternative sources for preparation
of Platelet Concentrate...
Platelet preparation by Buffy coat pooling method is
practiced in European countries, US, Latin America and
in other Asia...
 Even sometimes it is very difficult to get
donor because of stringent selection
procedure and people do not have time fo...
• Establish buffy coat pooling method to harvest platelet
equivalent to SDP.
- Improve platelet yield.
- Meet emergency re...
• Prepared 600 Pooled platelet units from 2400 units of blood
collected in Top and Bottom bags (Pooling of 4 buffy coat
ba...
• Collection of blood in Terumo penpol
Quadruple top and bottom bags.
• Process Quadruple top and bottom bags
in TACE.
• P...
• The Process of pooling Buffy coats can be performed in 2
different ways
• Train Method: Here the buffy coats are sterile...
Blood collected in Terumo Penpol 450ml TOP &
BOTTOM bag and kept in room temperature at
22*C for 2 hours.
Blood bags are c...
4 or 6 units of buffy from same group were connected
serially using TSCD.
Connected buffy coats with
TSCD
Connected buffy ...
Remove the pooled bag( bottom end) and
connect to IMUGARDIII PL filter integrated
with platelet storage bag using TSCD.
Po...
Pooling with BP Kit
(TERUFLEX Method)
BP Kit is integrated with Buffy coat pooling arm,
Platelet medium arm, Leukocyte fil...
TTI Screening
All four units for Buffy Coat Pooling are tested
for TTI markers by:
1. ELISA (4th
Generation)
2. ID-NAT (TM...
ID-NAT screened
ID-NAT screened
ID-NAT screened ID-NAT screened
Pooling
Platelet Concentrate
• Spin the pooled buffy at 1200 g for 9 minutes.Spin the pooled buffy at 1200 g for 9 minutes.
• After spinning express ou...
• Platelet yield.
• Platelet yield – Aphaeresis vs BC pooled.
• State of metabolism.
• Sterility test.
• Observational stu...
• Cell counter (Beckman coulter) – Platelet count and
WBC count is measured in Beckman Coulter Cell
Counter to study the p...
B.C.P.P. kept in platelet
agitator at 22O
C and
samples taken for
evaluation on each day up
to 5 days of storage to
analys...
The items shown in the table below were tested in order to
confirm the performance level of platelets.
Sl.
No
Test Content...
We had compared the QC
results of BC pooled PC and
Apheresis PC harvested by
HEMONETICS cell
separator.
Platelet count per bag is varying from
2.5 to 4.4 x 10¹¹ in 10 samples of
buffy coat pooled platelet. There was
no deterio...
We have checked the sterility
of pooled platelet prepared
after 5 hours and 24 hours of
preparation using Bact Alert
syste...
S
.no.
Parameter evaluated
1 Platelet count
2 pH
3 Lactic acid concentration
4 Glucose concentration
5 Pre and Post filtra...
Platelet Count and pH values Of Platelet prepared from Buffy Coat
Platelet count
(× 1011
)
Day 1 Day 3 Day 5 Day 6
3.31 3....
Comparison on platelet prepared from Buffy Coat vs
Aphaeresis (Average values)
Platelet count (× 1011
)
Buffy coat -PC Aph...
Viability & metabolic function of platelets were maintained during 5 days storage
as depicted by the glucose & lactate lev...
• To assess the clinical effect of the buffy coat
pooled platelets, permission was obtained from
the administrative author...
 Acute Leukemia on Chemotherapy ,
thrombocytopenia (n =10).
 Aplastic Anaemia undergoing labour(n=4).
 Dengue (n=125).
...
(Post-transfusion – Pretransfusion Platelet count)
(10⁹/1) Х Body Surface Area (m²) / number of
platelets transfused (10¹¹...
Estimated Total Blood Volume Х Platelet Count Increment
No. of Platelets Transfused
PPR(%) =
Diagnosis CCI -1 After 1-
2 hour
PPR %
after 1
-2 hour
PPR % AT 12 -24 Hours
Dengue
n=125
7.5-8 80-
85%
40 -50
Follow up w...
AIIMS
Clinicians
are liking to
use B.C.P.P.
Blood Bank is observing following
points
• Better result is obtained by BCPP
w...
Cost effectiveness
• Cost of buffy coat pooled platelet is
less than Rs 3000 /-.
• BC Pooled platelet reduce the cost
of p...
• Ready to use platelet to meet
emergency requirement.
• Reduce the cost of platelet therapy.
• High quality platelet.
• L...
• We are open for your valuable
suggestions to help poor patients in
case of their need.
• Can buffy coat pooled platelets...
Buffy coat
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Buffy coat pooling of platelets Blood Bank AIIMS New Delhi

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  • Over 25 years in Europe
  • Buffy coat

    1. 1. Dr. Kabita Chatterjee (MD) Faculty-In- Charge Blood Bank (Main Hospital) A.I.I.M.S., New Delhi – 110029.
    2. 2. 2 Countries where the Buffy Coat production method is predominantly used for the preparation of platelets derived from whole blood
    3. 3. • A layer of mixed white cells and platelets created by high speed centrifugation of whole blood • It is neutral or buff in colour • Contains: most of the white cells and platelets and ~ 10 % of RBC
    4. 4. PC FFP PRP method PC Buffy coat method PR P Plasma RBC PRP RBC Plasm a
    5. 5. 5
    6. 6. Platelets prepared through buffy method Activated Platelet prepared through PRP method PLATELET – Buffy coat V/s PRP Platelet Morphological Scoring
    7. 7.  In recent years, platelet rich buffy coat (BC) has become an alternative sources for preparation of Platelet Concentrate, particularly in Europe. It has been suggested that this method causes less platelet activation and damage during platelet preparation.
    8. 8. Platelet preparation by Buffy coat pooling method is practiced in European countries, US, Latin America and in other Asian counties.  A.I.I.M.S. is a reference center and getting platelet (SDP) request to correct thrombocytopenia from different clinical specialties. These patients are from different social status and most of them can not afford to purchase costly SDP kit (75% A.I.I.M.S. observation).
    9. 9.  Even sometimes it is very difficult to get donor because of stringent selection procedure and people do not have time for the procedure(1 ½ hours to 2 hours ). For BTS SDP is a routine procedure but for the clinicians sometimes they need urgent SDP to correct thrombocytopenia especially at the odd hours and holidays: Buffy coat pooled platelet is of great help in this situation(managing dengue crisis).
    10. 10. • Establish buffy coat pooling method to harvest platelet equivalent to SDP. - Improve platelet yield. - Meet emergency requirement for platelet. - Reduce cost to poor patients. - Reduce leukocyte contamination in platelet
    11. 11. • Prepared 600 Pooled platelet units from 2400 units of blood collected in Top and Bottom bags (Pooling of 4 buffy coat bags). • Conducted study on platelet yield, storage parameters and also on sterility parameters(n=125). • Observational study conducted on transfused patients (n= 100). • Platelet yield was compared with Aphaeresis platelet (n =25).
    12. 12. • Collection of blood in Terumo penpol Quadruple top and bottom bags. • Process Quadruple top and bottom bags in TACE. • Pooling of buffy coat. • Second separation & Filtration. • QC analysis. • Observational study on transfused patients. Methods
    13. 13. • The Process of pooling Buffy coats can be performed in 2 different ways • Train Method: Here the buffy coats are sterile docked to each other. All the buffy coat residues are pooled into one of the buffy coat bag, One unit Plasma is added to the same. Then the pooled unit is sterile docked to a platelet storage container with a leukoreduction filter and centrifuged. Our Study is based on this method. • BP Kit Method: refer to picture(TERUFLEX).
    14. 14. Blood collected in Terumo Penpol 450ml TOP & BOTTOM bag and kept in room temperature at 22*C for 2 hours. Blood bags are centrifuged at heavy spin and separated using TACE II
    15. 15. 4 or 6 units of buffy from same group were connected serially using TSCD. Connected buffy coats with TSCD Connected buffy coats with TSCD Pool 4 units and rinse with plasma Pool 4 units and rinse with plasma
    16. 16. Remove the pooled bag( bottom end) and connect to IMUGARDIII PL filter integrated with platelet storage bag using TSCD. Pooled bag IMUGUARD III PL storage bag integrated with filter
    17. 17. Pooling with BP Kit (TERUFLEX Method) BP Kit is integrated with Buffy coat pooling arm, Platelet medium arm, Leukocyte filter, Pooling bag and Platelet storage bag
    18. 18. TTI Screening All four units for Buffy Coat Pooling are tested for TTI markers by: 1. ELISA (4th Generation) 2. ID-NAT (TMA Technology) Buffy Coat pooling starts when these two reports are released by 5:00 PM We prefer to do polling by same blood group.
    19. 19. ID-NAT screened ID-NAT screened ID-NAT screened ID-NAT screened Pooling Platelet Concentrate
    20. 20. • Spin the pooled buffy at 1200 g for 9 minutes.Spin the pooled buffy at 1200 g for 9 minutes. • After spinning express out top layer which contains plateletAfter spinning express out top layer which contains platelet to Platelet storage bag through Leukocyte removal filterto Platelet storage bag through Leukocyte removal filter Final product- Platelets harvested from pooled Buffy
    21. 21. • Platelet yield. • Platelet yield – Aphaeresis vs BC pooled. • State of metabolism. • Sterility test. • Observational study. • Platelet increment.
    22. 22. • Cell counter (Beckman coulter) – Platelet count and WBC count is measured in Beckman Coulter Cell Counter to study the platelet count. • Blood gas analyzer (Nova) – PH, pCo2, pO2, glucose and lactate count is also taken during 5 days storage study. • Bact Alert(Biomerieux) to check Bacterial contamination.
    23. 23. B.C.P.P. kept in platelet agitator at 22O C and samples taken for evaluation on each day up to 5 days of storage to analyse the count and viability of the platelet.
    24. 24. The items shown in the table below were tested in order to confirm the performance level of platelets. Sl. No Test Content to be confirmed 1 Platelet Count State of Platelets during preservation 2 pH Preservation environment, State of metabolism 3 pCO2, Partial pressure 4 pO2 Partial pressure 5 Lactic Acid concentration State of metabolism 6 Glucose concentration State of metabolism
    25. 25. We had compared the QC results of BC pooled PC and Apheresis PC harvested by HEMONETICS cell separator.
    26. 26. Platelet count per bag is varying from 2.5 to 4.4 x 10¹¹ in 10 samples of buffy coat pooled platelet. There was no deterioration in the count during its 6 days storage period and meets the quality control requirements of Council of Europe guidelines for apheresis platelet.
    27. 27. We have checked the sterility of pooled platelet prepared after 5 hours and 24 hours of preparation using Bact Alert system. None of the pooled platelet showed the evidence of bacterial contamination at 5 hours and,24 hours.
    28. 28. S .no. Parameter evaluated 1 Platelet count 2 pH 3 Lactic acid concentration 4 Glucose concentration 5 Pre and Post filtration WBC count 6 Pre and Post filtration platelet count
    29. 29. Platelet Count and pH values Of Platelet prepared from Buffy Coat Platelet count (× 1011 ) Day 1 Day 3 Day 5 Day 6 3.31 3.36 3.37 3.36 pH 7.06 7.09 7.08 7.12  Platelet count was found to meet the Council of European guidelinesPlatelet count was found to meet the Council of European guidelines requirement No significant change inrequirement No significant change in pH during 5 days storageduring 5 days storage
    30. 30. Comparison on platelet prepared from Buffy Coat vs Aphaeresis (Average values) Platelet count (× 1011 ) Buffy coat -PC Aphaeresis-PC 3.31 3.21 pH 7.06 6.95  No significant difference in the Platelet count and pHNo significant difference in the Platelet count and pH between SDP & BC pooled platelet . Both are found equal inbetween SDP & BC pooled platelet . Both are found equal in quality point of view.quality point of view.
    31. 31. Viability & metabolic function of platelets were maintained during 5 days storage as depicted by the glucose & lactate level during storage. BC Pooled platelets Glucose (mg/dl) and Lactate (mmol/L)Concentration 1St day 3rd Day 5th Day Glucose lactate Glucose lactate Glucose lactate 353.4 11.25 335.1 12.69 319.1 14.34
    32. 32. • To assess the clinical effect of the buffy coat pooled platelets, permission was obtained from the administrative authorities of A.I.I.M.S. to prepare and issue Buffy-coat Pooled platelets to the patients (The matter is in knowledge of DCGI and TRG of NACO). • Patients who are receiving buffy coat pooled platelets will have to replace three units of blood. • By this policy A.I.I.M.S. blood bank indent is on rise.
    33. 33.  Acute Leukemia on Chemotherapy , thrombocytopenia (n =10).  Aplastic Anaemia undergoing labour(n=4).  Dengue (n=125).  GI Surgery ( n=4)  Replacement surgery in orthopaedics (n=10).  Misseleneous i.e.. DIC or Sepsis etc( n=5).
    34. 34. (Post-transfusion – Pretransfusion Platelet count) (10⁹/1) Х Body Surface Area (m²) / number of platelets transfused (10¹¹) Corrected count increment was calculated by the formula. CCI =
    35. 35. Estimated Total Blood Volume Х Platelet Count Increment No. of Platelets Transfused PPR(%) =
    36. 36. Diagnosis CCI -1 After 1- 2 hour PPR % after 1 -2 hour PPR % AT 12 -24 Hours Dengue n=125 7.5-8 80- 85% 40 -50 Follow up was not done in 50 patients due to time constraints, issue in emergency,short stay etc
    37. 37. AIIMS Clinicians are liking to use B.C.P.P. Blood Bank is observing following points • Better result is obtained by BCPP within 24 hours of collection • Our rigorous efforts to create a pool of voluntary aphaeresis donors has failed to gain momentum and this forced us to explore the usage of buffy coat pooled platelets as an alternative of SDP.
    38. 38. Cost effectiveness • Cost of buffy coat pooled platelet is less than Rs 3000 /-. • BC Pooled platelet reduce the cost of platelet therapy.
    39. 39. • Ready to use platelet to meet emergency requirement. • Reduce the cost of platelet therapy. • High quality platelet. • Leukodepleted platelet prevents WBC associated transfusion reactions. • Multiple donor exposure risk is reduced as it is tested with NAT. Advantages of buffy coat pooling
    40. 40. • We are open for your valuable suggestions to help poor patients in case of their need. • Can buffy coat pooled platelets be used as an subsidized alternative of SDP? • Blood Bank A.I.I.M.S., is doing more work on it to improve its usage. A.I.I.M.S., Blood Bank need your suggestions
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