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대한생식의학회지 제36권 제4호 2010년
초기 계대 인간 배아줄기세포의 해동 후 효율적인 배양 방법
서울대학교 의학연구원 인구의학연구소1, 삼진제약주식회사 중앙연구소2, 서울대학교 의과대학 산부인과학교실3
백진아1, 김희선1,3, 설혜원1, 서진1, 정주원1, 윤보애1, 박용빈2, 오선경1,3, 구승엽1,3, 김석현1,3, 최영민1,3*, 문신용1,3,
Efficient Culture Method for Early Passage hESCs after Thawing
Jin Ah Baek1, Hee Sun Kim1,3, Hye Won Seol1, Jin Seo1, Juwon Jung1, Bo Ae Yoon1, Yong Bin Park2, Sun Kyung Oh1,3, Seung-Yup Ku1,3, Seok Hyun Kim1,3, Young Min Choi1,3*, Shin Yong Moon1,3
1Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, 199-1, Dongsung Dong, Jongno Gu, Seoul, 110-810, Korea, 2Central Research Institute, Sam Jin Pharm. Co. Ltd., Hwasung 445-746, Korea, 3Department of Obstetrics and Gynecology, Seoul National University College of Medicine, 101 Daehang Ro, Jongno Gu, Seoul, 110-744, Korea
Objective: Human embryonic stem cells (hESCs) have the capacity to differentiate into all of the cell types and therefore hold promise for cell therapeutic applications. In order to utilize this important potential of hESCs, enhancement of currently used technologies for handling and manipulating the cells is required. The cryopreservation of hESC colonies was successfully performed using the vitrification and slow freezing-rapid thawing method. However, most of the hESC colonies were showed extremely spontaneous differentiation after freezing and thawing. In this study, we were performed to rapidly collect of early passage hESCs, which was thawed and had high rate of spontaneously differentiation of SNUhES11 cell line. Methods: Four days after plating, partially spontaneously differentiated parts of hESC colony were cut off using finely drawn-out dissecting pipette, which is mechanical separation method. Results: After separating of spontaneously differentiated cells, we observed that removed parts were recovered by undifferentiated cells. Furthermore, mechanical separation method was more efficient for hESCs expansion after thawing when we repeated this method. The recovery rate after removing differentiated parts of hESC colonies were 55.0%, 74.5%, and 71.1% when we have applied this method to three passages. Conclusion: Mechanical separation method is highly effective for rapidly collecting and large volumes of undifferentiated cells after thawing of cryopreserved early passage hESCs.
키워드 : Human embryonic stem cells (hESCs), Cryopreservation-Thawing, Early passage, Mechanical separation method
교신저자 : ymchoi@snu.ac.kr
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