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대한생식의학회지 제31권 제4호 2010년
양수 세포를 이용한 인간배아줄기세포의 배양
서울대학교 의과대학 산부인과학교실1, 의학연구원 인구의학연구소2
김희선1,2, 설혜원2,안희진2,오선경1,2,구승엽1,김석현1,최영민1,김정구1,문신용1,2
Human Amniotic Fluid Cells Support Expansion Culture of Human Embryonic Stem Cells
Hee Sun Kim1,2, Hye Won Seol2, Hee Jin Ahn2, Sun Kyung Oh1,2, Seung Yup Ku1,Seok Hyun Kim1, Young Min Choi1, Jung Gu Kim1, Shin Yong Moon1,2
1Department of Obstetrics and Gynecology, 2Institute of Reproductive Medicine and Population,Medical Research Center, College of Medicine, Seoul National University, Seoul, Korea
Objective: This study was performed to evaluate the possibility of prolonged culture of human embryonic stem cells (hESC; SNUhES2) on human amniotic fluid cells (hAFC), which had been storaged after karyotyping. Method: The hAFC was prepared for feeder layer in the presence of Chang's medium and STO medium (90% DMEM, 10% FBS) at 37℃ in a 5% CO2 in air atmosphere. Prior to use as a feeder layer, hAFC was mitotically inactivated by mitomycin C. The hESCs on hAFC were passaged mechanically every seven days with ES culture medium (80% DMEM/F12, 20% SR, bFGF). Results: The hAFC feeder layer support the growth of undifferentiated state of SNUhES2 for at least 59 passages thus far. SNUhES2 colonies on hAFC feeder appeared slightly angular and flatter shape as compared with circular and thicker colonies observed with STO feeder layer and showed higher level with complete undifferentiation in seven days. Like hESC cultured on STO feeders, SNUhES2 grown on hAFC expressed normal karyotype, positive for alkaline phosphatase activity, high telomerase activity, Oct-4, SSEA-3, SSEA-4, Tra-1-60 and Tra-1-81 and formed embryoid bodies (EBs). Conclusion: The hAFC supports undifferentiated growth of hESC. Therefore, these results may help to provide a clinically practicable method for expansion of hESC for cell therapies.
키워드 : Human embryonic stem cells (hESC), Human amniotic fluid cells (hAFC), Feeder layer, Undifferentiation, Expansion of hESC *위 연구는 과학기술부 21세기프로티어연구개발사업단인 세포응용연구사업단의 연구비 지원 (SC11011)에 의해 수행되었습니다.
교신저자 : shmoon@snu.ac.kr
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