Flow-cytometric screening of platelet antibodies with previously frozen cells.
In this study the flow-cytometric crossmatch results were compared between fresh cells and cells processed by various cryopreservation and storage methods. Platelets from healthy donors were incubated with 12 sera containing platelet reactive antibodies as well as with 62 control sera from blood donors. Direct comparisons were made between fresh platelets and platelets after freezing at -28 degrees C, -40 degrees C and -80 degrees C and in liquid nitrogen, using 6% DMSO as cryoprotectant. In addition, the effects of using controlled-rate freezing were evaluated. Finally we evaluated the application of the cryoprotectant ThromboSol. The best results were obtained after cryopreservation of the platelets with ThromboSol at -80 degrees C with controlled cooling rates. Using ThromboSol cryopreserved platelets, the sensitivity for the detection of incompatible platelets was 100% and the specificity was 97.1%, using the previous results obtained with flow-cytometry, MAIPA and LCT as a reference.


CONCLUSIONS
Platelets can be frozen using ThromboSol as the cryoprotectant, with controlled rate freezing and storage at -80 degrees C for the screening of platelet antibodies and for flow-cytometric crossmatch procedures. This system yields a reproducible and logistically simple method for platelet crossmatching that yields results superior to fresh cells and can be easily incorporated into standard clinical laboratory practices.
Döhlinger
Susanne
S
Department of Neurology, University of Göttingen, Göttingen, Germany.
Humpe
Andreas
A
Connor
Jerome
J
Köhler
Michael
M
Legler
Tobias J
TJ
D000906:Antibodies / Q000097:blood; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D001792:Blood Platelets / Q000276:immunology*; D015925:Cryopreservation / Q000592:standards*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D017713:Platelet Transfusion / Q000592:standards*; D015290:Second Messenger Systems
D000906:Antibodies; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide