Study On The IVM,IVF Of Oocytes And IVC Of Embryos In Ovine

Abstract: In this study, the aim is to optimize the systems of IVM, IVF of oocytes and IVC of embryos in ovine and further promote embryo biology engineer development. For this goal, ovine oocytes from diameter 2-6 mm surface follicles of ovaries from a slaughterhouse were matured in the base maturation medium TCM199 supplemented with 10% FBS to study the effects of FSH/LH concentrations, animal ages, seasons, the preservation temperatures of ovaries during transporting on IVM of ovine oocytes; the matured oocytes and ovine freezing sperms were fertilized in vitro to study the effects of fertilization systems, the handling methods of matured oocytes before fertilization on IVF of ovine oocytes; the matured ovine oocytes were activated in 5 μmol/L A23187 and 2 mmol/L 6-DMAP, and the embryos were cultured in the base culture medium mSOF to study the effects of amino acids and cysteamine on in vitro development of ovine embryos. The major results obtained are as follows.1. FSH/LH could effectively promote IVM of ovine oocytes and the suitable adding concentration was very important; in this study, the proper concentration was 10 μg/ml.2. The maturation rates of adult ovine oocytes were higher than that of prepubertal ones, but there was no significant difference between them. The NO.of collection ovine oocytes was more in estrous season than that in no-estrous season, but there was no significant difference between them; the ovine oocytes of estrous season was more easier matured in vitro than that ofno-estrous season.3. In this study, the suitable preservation temperature of ovine ovaries during transporting was 28-32 ℃.4. BO fertilization system was more suitable for the cleavage of ovine oocytes than TALP fertilization system. In this study, the denudation of the majority of expanded cumulus cells was benefical for cleavage .5. Amino acids could promote in vitro development of ovine embryos; but when added during 0~24-hour culture, it inhibited the cleavage of ovine oocytes; when added during culture after 24-hour, it could more promote in vitro development of ovine embryos than when added during the whole culture. When added EAA to culture medium during 24~72h culture, it could promote in vitro development of ovine embryos and the proper adding concentration was 1 ×.6. Cysteamine could enhance in vitro development of ovine embryos and the suitable adding concentration was very important; the proper concentration was 100 μmol/L.