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대한생식의학회지 제21권 제2호 2010년
체외성숙배양 조건이 마우스 난자의 체외수정 및 다정자침입에 미치는 영향
고려대학교 자연자원대학 응용동물과학과;고려대학교 자연자원대학 응용동물과학과;대구대학교 농과대학 축산학과;
박기상;이상호;송해범;,
In Vitro Fertilization and Polyspermy in Follicular Oocytes Matured in Various Culture Conditions
Park, Kee-Sang;Lee, Sang-Ho;Song, Hai-Bum;
Department of Animal Science, College of Natural Resources, Korea University;Department of Animal Science, College of Natural Resources, Korea University;Department of Animal Science, College of Agriculture, Teagu University;
ICR female mice aged 3 to 4 weeks, were stimulated with 7.5 IU PMS injection. At 48-52h post-PMS injection, ovaries were dissected out and oocytes-cumulus complexes(OCCs) were divided into three groups, cumulus-free oocytes(O), cumulus-free oocyte cocultured with cumulus cells(O+C) and OCC. The oocyte were cultured in TCM199 containing various protein sources, FCS, BSA or PVP with gonadotropins(Gns) for 24h. Spermatozoa were collected from cauda epididymis and capacitated in T6 + BSA for 2h. After oocyte maturation in vitro(IVM) in different experimental groups, matured oocytes were inseminated with the capacitated spermatozoa in T6 + BSA for 6h. In the groups of IVM in TCM + BSA or PVP, fertilization(IVF) did not occur efficiently. However, increased fertilization was found in TCM+ FCS group. The oocytes groups, with cumulus cells showed decreased polyspermy in FCS group (O; 31.8 %, O + C; 12.2 %, OCC; 16%), the addition of Gns did not prevent polyspermy in all three groups. The rates of fertilization increased in zona-free oocytes in PVP group. This results showed that culture system for IVM and IVF could be improved. Furthermore, PVP can be used for the substitution of protein source during maturation, and its low rate of fertilization has been found due to zona hardening which occurred in FCS-free medium.
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