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Comparison of platelet count and functional recovery after thawing and freezing according to cryopreservation methods Dump freezing vs controlled rate freezing



Comparison of platelet count and functional recovery after thawing and freezing according to cryopreservation methods Dump freezing vs controlled rate freezing



Blood 102(11): 138b



Introduction: The limit and the optimal method of the cryopreservation of platelets have not been determined. Moreover, the functional changes platelets after cryopreservation were not clearly defined. This study was conducted to determine the reasonable duration of platelet cryopreservation to maintain the optimal cell counts and function according to the method of cryopreservation method. Methods: We compared the recovery, expression of membrane GpIb, GpIIb/IIIa, and aggregatory function as well as cell counts of the platelets preserved in two different conditions(-80degreeC dump freezing, controlled rate freezing with -196degreeC liquid nitrogen storage). Total fourteen platelet samples from 4 healthy volunteer donors by apheresis and 10 platelet concentrates (PCs) by random blood donation were used for this study. Apheresis platelets were used for the measurement of functional recovery, PCs were used for the measurement of platelet count recovery. For cryopreservation, platelets with 5% DMSO were either inserted directly in -80degreeC freezer or in liquid nitrogen after computer-controlled rate freezing. Apheresis platelets were thawed after cryopreservation of 1 week, 2 weeks, 3 weeks, 4 weeks, and 12 weeks,and in-vitro function of thawed platelets were compared to those of platelets stored in 22degreeC liquid storage for 5 days(maximal storage duration). PCs were thawed after cryopreservation of 1 month, 3, 6, 9, 12, 15, 18 months, and cell recovery rate of thawed platelets were compared to those of initial platelet count before cryopreservation. Results: Platelets preserved at 22degreeC or cryopreserved with each condition displayed equivalent recovery (90%). With each cryopreservation procedures, platelets showed moderate loss of GpIb and retained more than 90% of GpIIb/IIIa in comparison with fresh platelets. At the third week, loss of GpIb in the directly frozen platelets was augmented compared with those of controlled rate frozen group. The aggregatory response to ristocetin of cryopreserved platelets showed no significant difference to platelets stored in 22degreeC liquid storage for 5 days. And the aggregatory function of platelet stored in -80degreeC dump freezing were comparable to that of platelet stored in controlled rate freezing with -196degreeC liquid nitrogen storage. However, controlled rate frozen platelets retained more aggregatory response to ristocetin and surface GpIb expression than those of directly frozen platelets at the 3rd, 4th, 12th week of storage. To 18 monthes of cryopreservation, platelet recovery rates after thawing were more than 80% in each group without differences(dump freezing:: 84%, controlled rate freezing: 89%) Conclusion: This study showed the possibility of preservation of in vitro functions of frozen-thawed platelets after 12 weeks of storage comparable to those of the liquid stored 5-day old platelets. And platelet recovery of thawed platelet after 18 monthes of cryopreservation was more than 80%. Lastly, cell count and functional recovery after thawing of the platelet stored in -80degreeC dump freezing were not signicantly different to those of the platelet stored in controlled rate freezing with liquid nitrogen storage.

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