Preparation of leukodepleted platelet concentrates from pooled buffy coats: prestorage filtration with Autostop BC.
OBJECTIVE
Our requirements for leukocyte-depleted platelet concentrates (LD-PC) for an adult patient are: platelets >240x10(9), leukocytes <5x10(6), volume of 150-400 ml; and at the end of storage a pH between 6.8 and 7.4 and presence of the swirling effect. Our aim was to develop a standardized, semiautomated method for the production of LD-PC, by pooling of buffy coats (BC), and prestorage leukoreduction by filtration.


METHODS
Whole blood was collected in Top and Bottom systems, and separated automatically with the Compomattrade mark G3 equipment into a red cell concentrate, a plasma and a BC. Subsequently, a pool of 5 BC was made, and 200 g plasma from one of the donors was added. Then, after soft spin centrifugation, the platelet rich plasma was leukocyte depleted by filtration using the Autostoptrade markBC filter, and stored in a 1,000 ml polyolefin platelet storage bag.


RESULTS
BC (n = 60) had a volume of 51+/-2 ml (mean +/- SD) with a hematocrit of 0.44+/-0.03 l/l and contained 80+/-5% of the platelets and 74+/-12% of the leukocytes of the whole blood. Routinely prepared LD-PC (n = 15,037) contained a median of 341x10(9) platelets (range 49-599x10(9)), with only 104/15,037 (0.7%) containing fewer than 240x10(9) platelets; the median volume was 263 ml (range 134-373 ml). In 118/917 (13%) LD-PC leukocytes were observed in the Nageotte hemocytometer, but only twice exceeding 1x10(6) leukocytes per unit, and none exceeding 5x10(6) (median <0. 6x10(6); range <0.6-1.41x10(6)). Storage experiments of the LD-PC (n = 12) revealed adequate oxygenation and maintenance of pH and swirling effect up to 9 days.


CONCLUSIONS
This method warrants with 99% confidence that LD-PC contain more than 240x10(9) platelets; with 97.5% confidence that 100% of the LD-PC contain <5x10(6) leukocytes, and with 95% confidence that more than 99% of the LD-PC contain fewer than 1x10(6) leukocytes; these LD-PC can be stored satisfactorily for up to 9 days.
Red Cross Blood Bank, Amsterdam, Amsterdam, The Netherlands. Blbadam@compuserve.com
R N
RN
Pietersz
P F
PF
van der Meer
I
I
Steneker
B
B
Hinloopen
W J
WJ
Dekker
A P
AP
van Zanten
H W
HW
Reesink
D001331:Automation; D001792:Blood Platelets / Q000166:cytology*; D002469:Cell Separation / Q000379:methods*; D002498:Centrifugation; D005374:Filtration / Q000295:instrumentation; D006801:Humans; D007937:Leukapheresis; D007962:Leukocytes / Q000166:cytology*; D013210:Staphylococcus / Q000302:isolation & purification