Korean J Hematol 2003; 38(3):
Published online September 30, 2003
© The Korean Society of Hematology
혈소판의 냉동보존 방법의 차이에 따른 활성도의 변화
박경화, 김병수, 배숙영, 윤수영, 설혜령, 최종권, 성화정, 윤소영, 최인근, 오상철, 서재홍, 최철원, 신상원, 김열홍, 김준석
고려대학교 의과대학 혈액종양내과,
고려대학교 의과대학 진단검사의학과
Comparison of the Activities of Platelets with Different Cryopreservation Methods
Background:
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 limit and optimal method for cryopreservation of platelet concentrates.
Methods:
We compared the recovery, expression of membrane Gplb, Gpllb/llla, and aggregatory function of the platelets preserved in three different conditions. Platelet samples were collected from four healthy volunteer donors by apheresis, and placed in 22℃ agitator for standard preservation. For cryopreservation, after treating 5% DMSO, platelets were either inserted directly in -80℃ freezer or in liquid nitrogen after computer-controlled rate freezing. After storage for 5 days, 1 week, 2 weeks, 3 weeks, 4 weeks, and 12 weeks, platelets were thawed and analyzed for the evaluation of in vitro functions.
Results:
Platelets preserved at 22℃ or cryopreserved with each condition displayed equivalent recovery (90%). With each cryopreservation procedures, platelets showed moderate loss of Gplb and retained more than 90% of Gpllb/llla in comparison with fresh platelets. At the third week, loss of Gplb in the directly frozen platelets was augmented compared with those of controlled rate frozen group. The aggregatory response to ristocetin began to decrease at 12 weeks of storage. However, controlled rate frozen platelets retained more aggregatory response to ristocetin and surface Gplb expression than those of directly frozen platelets at 3, 4, 12 weeks of storage.
Conclusion:
This study showed the possibility of moderate preservation of in vitro functions of frozen-thawed platelets after 12 weeks of storage compared with those of the liquid stored 5-days old platelets.
Keywords Platelets; Cryopreservation; Membrane glycoprotein
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Korean J Hematol 2003; 38(3): 169-175
Published online September 30, 2003
Copyright © The Korean Society of Hematology.
혈소판의 냉동보존 방법의 차이에 따른 활성도의 변화
박경화, 김병수, 배숙영, 윤수영, 설혜령, 최종권, 성화정, 윤소영, 최인근, 오상철, 서재홍, 최철원, 신상원, 김열홍, 김준석
고려대학교 의과대학 혈액종양내과,
고려대학교 의과대학 진단검사의학과
Comparison of the Activities of Platelets with Different Cryopreservation Methods
Kyong Hwa Park, Byung Soo Kim, Sook Young Bae, Soo Young Yoon, Hye Ryoung Sul, Jong Gwon Choi, Hwa Jung Sung, So Young Yoon, In Keun Choi, Sang Cheul Oh, Jae Hong Seo, Chul Won Choi, Sang Won Shin, Yeul Hong Kim, Jun Seok Kim
Division of Hemato Oncology, Department of Internal Medicine and Laboratory Medicine, Korea University College of Medicine, Seoul, Korea
Abstract
Background:
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 limit and optimal method for cryopreservation of platelet concentrates.
Methods:
We compared the recovery, expression of membrane Gplb, Gpllb/llla, and aggregatory function of the platelets preserved in three different conditions. Platelet samples were collected from four healthy volunteer donors by apheresis, and placed in 22℃ agitator for standard preservation. For cryopreservation, after treating 5% DMSO, platelets were either inserted directly in -80℃ freezer or in liquid nitrogen after computer-controlled rate freezing. After storage for 5 days, 1 week, 2 weeks, 3 weeks, 4 weeks, and 12 weeks, platelets were thawed and analyzed for the evaluation of in vitro functions.
Results:
Platelets preserved at 22℃ or cryopreserved with each condition displayed equivalent recovery (90%). With each cryopreservation procedures, platelets showed moderate loss of Gplb and retained more than 90% of Gpllb/llla in comparison with fresh platelets. At the third week, loss of Gplb in the directly frozen platelets was augmented compared with those of controlled rate frozen group. The aggregatory response to ristocetin began to decrease at 12 weeks of storage. However, controlled rate frozen platelets retained more aggregatory response to ristocetin and surface Gplb expression than those of directly frozen platelets at 3, 4, 12 weeks of storage.
Conclusion:
This study showed the possibility of moderate preservation of in vitro functions of frozen-thawed platelets after 12 weeks of storage compared with those of the liquid stored 5-days old platelets.
Keywords: Platelets, Cryopreservation, Membrane glycoprotein
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