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대한생식의학회지 제27권 제4호 2010년
생쥐배아의 부화와 탈각에 미치는 Pronase의 영향
서울대학교 의과대학 산부인과학교실1, 서울대학교 의학연구원 인구의학연구소2, 성균관대학교 의과대학 삼성제일병원 산부인과3
문신용1,2, 최성미1,2, 김희선1,2, 류범용1,2, 오선경1,2, 서창석1, 김석현1, 최영민1, 김정구1, 최규홍2,3, 이진용1,
Effect of Pronase Treatment on Mouse Embryos: Improving Hatching and Hatched Rates
Shin Young Moon1,2, Sung Mi Choi1,2, Hee Sun Kim1,2, Buom Yong Ryu1,2, Sun Kkyng Oh1,2, Chang Suk Suh1, Seok Hyun Kim1, Young Min Choi1, Jung Gu Kim1, Kyu Hong Choi2,3, Jin Yong Lee1
Department of Obstetrics and Gynecology, College of Medicine1, Institute of Reproductive Medicine and Population, Medical Research Center2, Seoul National University, Department of Obstetrics and Gynecology, Samsung Cheil Hospital, Sungkyunkwan University School of medicine3, Seoul, Korea
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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 F1 animals. Fresh and cryo-thawed morula embryos were exposed to 0.5, 1.0, 2.0, 5.0 ug/ml pronase in Ham's F10 for 72hrs. 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 ug/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.
키워드 : Pronase, Hatching, Assisted hatching
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