Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
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Effects of Pronuclear Age in Freezing of Mouse Embryos on Survival and Development in Vitro after Cryopreservation

동결보존시 생쥐 전핵배아의 시기에 따른 생존율과 발생율의 비교

  • Kim, H.S. (Institute of Reproductive Medicine and Population, Medical Research Center College of Medicine, Seoul National University) ;
  • Ryu, B.Y. (Institute of Reproductive Medicine and Population, Medical Research Center College of Medicine, Seoul National University) ;
  • Oh, S.K. (Institute of Reproductive Medicine and Population, Medical Research Center College of Medicine, Seoul National University) ;
  • Suh, C.S. (Department of Obstetrics and Gynecololgy, College of Medicine, Seoul National University) ;
  • Kim, S.H. (Department of Obstetrics and Gynecololgy, College of Medicine, Seoul National University) ;
  • Choi, Y.M. (Department of Obstetrics and Gynecololgy, College of Medicine, Seoul National University) ;
  • Kim, J.G. (Department of Obstetrics and Gynecololgy, College of Medicine, Seoul National University) ;
  • Moon, S.Y. (Department of Obstetrics and Gynecololgy, College of Medicine, Seoul National University) ;
  • Lee, J.Y. (Department of Obstetrics and Gynecololgy, College of Medicine, Seoul National University)
  • 김희선 (서울대학교 의학연구원 인구의학연구소) ;
  • 류범용 (서울대학교 의학연구원 인구의학연구소) ;
  • 오선경 (서울대학교 의학연구원 인구의학연구소) ;
  • 서창석 (서울대학교 의과대학 산부인과학교실) ;
  • 김석현 (서울대학교 의과대학 산부인과학교실) ;
  • 최영민 (서울대학교 의과대학 산부인과학교실) ;
  • 김정구 (서울대학교 의과대학 산부인과학교실) ;
  • 문신용 (서울대학교 의과대학 산부인과학교실) ;
  • 이진용 (서울대학교 의과대학 산부인과학교실)
  • Published : 1998.03.30
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Abstract

This study was designed to evaluate the influence of pronuclear age on the survival and post-thawing development after cryopreservation of mouse embryos. Freezing and thawing were performed in the different pronuclear stages of mouse embryos after IVF. Embryos were obtained from $F_1$ hybrid mice and classified into 4 groups according to the pronuclear stage (6hr, 9hr, 12hr and 15hr after insemination). Pronuclear ova were slowly cooled in a biological freezer using 1.5M 1,2-propanediol and 0.1M sucrose as cryoprotectant. Thawing was done at room temperature and 1,2-propanediol was removed by multi-step dilutions. Both frozen-thawed embryos and control fresh embryos were cultured in vitro in Ham's F-10 medium supplemented with 4mg/ml BSA. In control group, the development rate after 48hr was 99.3%, and the complete hatching rate after 144hr was 61.3%. In experimental groups, the survival rate after thawing was 95.4% in 6hr, 88.7% in 9hr, 75.2% in 12hr and 62.4% in 15hr after insemination, the development rate after 48hr was 61.1, 77.0, 67.0 and 79.6%, respectively, and the complete hatching rate after 144hr was 25.7, 43.7, 42.2 and 60.0%, respectively. The survival rate in 15hr was significantly lower (p<0.05) compared with other groups. In vitro development rates after 48hr were similar in all groups, but complement hatching rate was significantly lower (p<0.05) in 6hr group. In conclusion, cryopreservation of mouse pronuclear ova with 2 distinct pronuclei (9hr and 12hr groups) showed better results after thawing compared with early (6hr group) or late pronuclear ova just prior to cleavage (15hr group).

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