Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
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Human Amniotic Fluid Cells Support Expansion Culture of Human Embryonic Stem Cells

양수 세포를 이용한 인간배아줄기세포의 배양

  • Kim, Hee-Sun (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University) ;
  • Seol, Hye-Won (Institute of Reproductive Medicine and Population, Medical Research Center, College of Medicine, Seoul National University) ;
  • Ahn, Hee-Jin (Institute of Reproductive Medicine and Population, Medical Research Center, College of Medicine, Seoul National University) ;
  • Oh, Sun-Kyung (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University) ;
  • Ku, Seung-Yup (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University) ;
  • Kim, Seok-Hyun (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University) ;
  • Choi, Young-Min (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University) ;
  • Kim, Jung-Gu (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University) ;
  • Moon, Shin-Yong (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University)
  • 김희선 (서울대학교 의과대학 산부인과학교실) ;
  • 설혜원 (서울대학교 의과대학 의학연구원 인구의학연구소) ;
  • 안희진 (서울대학교 의과대학 의학연구원 인구의학연구소) ;
  • 오선경 (서울대학교 의과대학 산부인과학교실) ;
  • 구승엽 (서울대학교 의과대학 산부인과학교실) ;
  • 김석현 (서울대학교 의과대학 산부인과학교실) ;
  • 최영민 (서울대학교 의과대학 산부인과학교실) ;
  • 김정구 (서울대학교 의과대학 산부인과학교실) ;
  • 문신용 (서울대학교 의과대학 산부인과학교실)
  • Published : 2004.12.30
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Abstract

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 $37^{circ}C$ in a 5% $CO_2$ 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.

Keywords

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