Effect of Pronase Treatment on Mouse Embryos: Improving Hatching and Hatched Rates

생쥐배아의 부화와 탈각에 미치는 Pronase의 영향

  • Moon, Shin-Yong (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University, Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University) ;
  • Choi, Sung-Mi (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University, Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University) ;
  • Kim, Hee-Sun (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University, Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University) ;
  • Ryu, Buom-Yong (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University, Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University) ;
  • Oh, Sun-Kyung (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University, Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University) ;
  • Suh, Chang-Suk (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) ;
  • Choi, Kyu-Hong (Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Department of Obstetrics and Gynecology, Samsung Cheil Hospital, Sungkyunkwan University School of Medicine) ;
  • Lee, Jin-Yong (Department of Obstetrics and Gynecology, College of Medicine, Seoul National University)
  • 문신용 (서울대학교 의과대학 산부인과학교실, 서울대학교 의학연구원 인구의학연구소) ;
  • 최성미 (서울대학교 의과대학 산부인과학교실, 서울대학교 의학연구원 인구의학연구소) ;
  • 김희선 (서울대학교 의과대학 산부인과학교실, 서울대학교 의학연구원 인구의학연구소) ;
  • 류범용 (서울대학교 의과대학 산부인과학교실, 서울대학교 의학연구원 인구의학연구소) ;
  • 오선경 (서울대학교 의과대학 산부인과학교실, 서울대학교 의학연구원 인구의학연구소) ;
  • 서창석 (서울대학교 의과대학 산부인과학교실) ;
  • 김석현 (서울대학교 의과대학 산부인과학교실) ;
  • 최영민 (서울대학교 의과대학 산부인과학교실) ;
  • 김정구 (서울대학교 의과대학 산부인과학교실) ;
  • 최규홍 (서울대학교 의학연구원 인구의학연구소, 성균관대학교 의과대학 삼성제일병원 산부인과) ;
  • 이진용 (서울대학교 의과대학 산부인과학교실)
  • Published : 2000.12.30

Abstract

Objective: Hatching of the blastocyst from the zona pellucida (ZP) is a key event in mammalian implantation. In vivo, two factors have been identified as possible mediators of hatching: lysis of the ZP by substances elaborated either from the embryo or female reproductive tract and pressure exerted on the zona by expansion of the blastocyst. Two methods of zona manipulation were already in use to enhance the ability of embryos to hatch: mechanical PZD and chemical ZD by acidic Tyrode's solution. But several controversies of each method have been reported. The purpose of this study was to investigate the effect of pronase for mouse embryo hatching. Methods: Mouse embryos were obtained following ovulation induction of $F_1$ animals. Fresh and cryo-thawed morula embryos were exposed to 0.5, 1.0, 2.0, 5.0 ${\mu}g/ml$ pronase in Ham's F10 for 72 hrs. Main outcome measures were the rates of partial hatching and completely hatched blastocysts, and cell number of it. Results: In fresh and cryo-thawed group, the rates of completely hatched blastocyst were significantly higher in 5 ${\mu}g/ml$ pronase treatment group than control group. There was no difference in completely hatched blastocyst total cell number between pronase treatment group and control group. This suggest that pronase treatment did not harmful in mouse embryo development. In pronase treatment group, zona pellucida were thinner than control group. Conclusion: The addition of pronase to culture media may accelerate the hatching of embryo. So, enzymatic treatment of the zona may provide a valuable and effective assisted hatching technique for human in-vitro fertilization-embryo transfer.

Keywords