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Journal of the Korean Pediatric Society 2000;43(12):1576-1582.
The Changes of Apoptosis and Viability in Stem Cells after Thawing with Lapse of the Time
Seun Joo Park1, Sang Young Bae1, Kyung Ha Ryu1, Il Tae Whang1, Kyung Hyo Kim1, Young Mi Hong1, Kyung Hee Kim1, Keun Lee1, Joo Young Seoh2
1Department of Pediatrics, College of Medicine, Ewha Womans University, Seoul, Korea
2Department of Microbiology, College of Medicine, Ewha Womans University, Seoul, Korea
조혈세포 해동 후 시간 경과에 따른 세포 고사 및 세포 생존력 변화에 관한 연구
박선주1, 배상영1, 유경하1, 황일태1, 김경효1, 홍영미1, 김경희1, 이 근1, 서주영2
1이화여자대학교 의과대학 소아과학교실
2이화여자대학교 의과대학 미생물학교실
Abstract
Purpose
: Thawed stem cells should be infused as early as possible because delay of infusion leads to decrease of cell viability and formation of DNA clumping. The procedure of 10% Dimethyl sulfoxide(DMSO) removal and a long distance from the thawing location to the patient are the main causes of delay of infusion that results in the loss of cell viability and apoptosis. Therefore, we investigated the changes of cell viability and apoptosis after thawing with lapse of time.
Methods
: Five samples of mobilized peripheral blood were evaluated. We measured cell viability, colony forming unit(CFU) and apoptosis at 0, 1, 2, 4, and 24 hours after thawing. The state of stem cells were divided into live, apoptotic and dead with double staining using annexin V and 7-aminoactinomycin D(7-AAD) in flow cytometry.
Results
: Viability of the total cells after thawing was 77.3(53.3-97.7)%. The percentage which recovered to initial CFU at 1, 2, 4 and 24 hours after thawing decreased to 63.9%, 50.2%, 45.8% and 11.6%, respectively. The proportion of apoptotic cells among CD34+ cells after thawing were increased from 0.2% at 0 hour to 16.5% at 1 hour, 21.9% at 2 hours, and then decreased to 15.0% at 4 hours, 2.7% at 24 hours because they were replaced by dead cells.
Conclusion
: Thawed cells changed to apoptotic and had less colony forming capacity from 1 hour after thawing, and were then replaced by dead cells from 4 hours after thawing.
Key Words: Thawing, Cell viability, Apoptosis, Time, CFU


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