Development and Reproduction (한국발생생물학회지:발생과생식)

The Korean Society of Developmental Biology (한국발생생물학회)
권호 표지

Improving the Survival and Maintenance of the Undifferentiated State of Cryopreserved Human Embryonic Stem Cells by Extended Incubation with Feeder Cells Overnight before Vitrification

동결에 앞서 시행된 지지세포와의 추가 공배양이 인간 배아줄기세포의 유리화 동결보존 후 생존율과 미분화 유지에 미치는 영향

  • Cha, Soo-Kyung (Fertility Center of CHA General Hospital, CHA Research Institute, Pochon CHA University College of Medicine) ;
  • Choi, Kyoung-Hee (Fertility Center of CHA General Hospital, CHA Research Institute, Pochon CHA University College of Medicine) ;
  • Shin, Ju-Mi (Fertility Center of CHA General Hospital, CHA Research Institute, Pochon CHA University College of Medicine) ;
  • Park, Kyu-Hyung (CHA Stem Cell Institute, Pochon CHA University College of Medicine) ;
  • Yoon, Tae-Ki (Fertility Center of CHA General Hospital, CHA Research Institute, Pochon CHA University College of MedicineCHA Stem Cell Institute, Pochon CHA University College of Medicine) ;
  • Chung, Hyung-Min (CHA Stem Cell Institute, Pochon CHA University College of Medicine) ;
  • Lee, Dong-Ryul (Fertility Center of CHA General Hospital, CHA Research Institute, Pochon CHA University College of Medicine)
  • 차수경 (포천중문의과대학교 차병원 여성의학연구소) ;
  • 최경희 (포천중문의과대학교 차병원 여성의학연구소) ;
  • 신주미 (포천중문의과대학교 차병원 여성의학연구소) ;
  • 박규형 (포천중문의과대학교 줄기세포연구소) ;
  • 윤태기 (포천중문의과대학교 차병원 여성의학연구소) ;
  • 정형민 (포천중문의과대학교 줄기세포연구소) ;
  • 이동률 (포천중문의과대학교 차병원 여성의학연구소)
  • Published : 2008.09.30
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Abstract

This study was conducted to develop an efficient cryopreservation method of human embryonic stem (ES) cells using vitrification. In an initial experiment, sub-clumps of human ES cells (CHA-hES3 and CHA-hES4) were vitrified using grids after incubation with STO feeder cells for 1 or 16 h (Groups 1-1 and 1-2, respectively). After storage for $2{\sim}4$ months, thawed clumps were re-plated on a fresh feeder layer. The survival rates of warmed CHA-hES3 and CHA-hES4 cells of Group 1-2 were significantly higher than those of the corresponding Group 1-1 cells. In the second experiment, human ES cells were vitrified after incubation with feeder or feeder-conditioned medium (Groups 2-1 to -7). Relative mRNA expression of BM proteins and survival rates were increased following incubation of ES cells with fresh feeder cells for 16 h. In conclusion, increasing of tight adhesion between ES cells by extended incubation with feeder could reduce cryoinjury after vitrifying/warming.

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