Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
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The Study on Vitrification and Ultrarapid Thawing of Human Embryonic Stem Cells

인간 배아 줄기세포의 초자화 동결 및 초급속 융해에 관한 연구

  • Moon, Shin-Yong (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University) ;
  • Park, Yong-Bin (Institute of Reproductive Medicine and Population, Medical Research Center) ;
  • Kim, Hee-Sun (Institute of Reproductive Medicine and Population, Medical Research Center) ;
  • Sung, Ki-Chung (Institute of Reproductive Medicine and Population, Medical Research Center) ;
  • Oh, Sun-Kyung (Institute of Reproductive Medicine and Population, Medical Research Center) ;
  • Chun, Dae-Woo (Institute of Reproductive Medicine and Population, Medical Research Center) ;
  • Suh, Chang-Suk (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) ;
  • Lee, Jin-Yong (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)
  • 문신용 (서울대학교 의과대학 산부인과학교실) ;
  • 박용빈 (의학연구원 인구의학연구소) ;
  • 김희선 (의학연구원 인구의학연구소) ;
  • 성기청 (의학연구원 인구의학연구소) ;
  • 오선경 (의학연구원 인구의학연구소) ;
  • 천대우 (의학연구원 인구의학연구소) ;
  • 서창석 (서울대학교 의과대학 산부인과학교실) ;
  • 최영민 (서울대학교 의과대학 산부인과학교실) ;
  • 김정구 (서울대학교 의과대학 산부인과학교실) ;
  • 이진용 (서울대학교 의과대학 산부인과학교실) ;
  • 김석현 (서울대학교 의과대학 산부인과학교실)
  • Published : 2002.03.30
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Abstract

Objective: This study was carried out to establish the effectiveness of the vitrification method and the optimal cryoprotectants in the cryopreservation of human embryonic stem cells (ESC). Materials and Methods: Human ESC clumps established at Seoul National University Hospital (SNUhES 1) were cryopreserved with the vitrification method using the EM grid. EDS and EFS40 were used as vitrification solutions. Results: Between the EDS and EFS40 groups, there was no significant difference in the recovery rate after cryopreservation of human ESC. The formation rates of ESC colonies in the vitrified groups were significantly lower than those in the control ESC group (p<0.05, p<0.05). In addition, the formation rate of ESC colonies in the EDS group was significantly higher than that in the EFS40 group (p<0.05). The ESC colonies in the vitrified groups were significantly smaller after culture duration of 2 and 4 days, respectively, compared with the control ESC group (p<0.1, p<0.05). However, these effects could be reduced to nonsignificant level by the additional culture of ESC colonies. The vitrified human ESC retained the properties of pluripotent cells, including the expression of cell surface. markers for the undifferentiated cells such as alkaline phosphatase and SSEA-4 (stage-specific embryonic antigen-4), and the expression of transcription factor Oct-4 (octamer-binding transcription factor-4), and the normal karyotype. Conclusion: The vitrification method using the EM grid and EDS solution was confirmed to be very effective for the cryopreservation of human ESC.

Keywords

References

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