摘要
[目的]研究孕马血清促性腺激素(PMSG;eCG)对狗卵母细胞体外成熟效果的影响。[方法]采用切割法收集卵巢表面卵丘卵母细胞复合体(COCs),并且随机分为4组,在添加不同浓度的PMSG的基础培养液(含有10%胎牛血清的DMEM)中,5%CO2,38.5℃培养箱内培养48 h。在含有0.1%透明质酸酶的D-PBS中用细小玻璃管剥离卵丘细胞从而获得裸卵,为了鉴别核成熟期,用4%甲醛溶液固定裸卵15 min,然后用10μg/ml Hoechst33342染色、压片,荧光显微镜下观察核形态。[结果]体外培养48 h后,添加PMSG处理组卵丘扩散明显,但发育到MⅠ-MⅡ的比率都没有显著差异(P(0.05)。100 IU/ml PMSG处理组GVBD期率显著低于对照组。[结论]添加PMSG改善了卵丘扩散的效果,但是没有提高核成熟率,需进一步改进狗卵母细胞体外成熟率的培养体系。
[ Objective ] The aim of this study was to evaluate the effects of PMSG on nuclear maturation of canine oocyte in vitro. [ Method ] Cumulus-ooeyte complexes (COCs) were released by slicing. They were randomly divided into four groups and cultured in DMEM containing 10% foetal bovine serum (FBS) supplemented with different concentrations of PMSG at 38.5 ℃, 5% CO2 in air for 48 h. To evaluate the nuclear meiotic stage, oocytes were denuded by placing them in D-PBS supplemented with 0.1% hyaluronidase, and then gently pipetted using small glass pipettes. Denuded oocytes were fixed with 4% formaldehyde for 15 rain and 10 μg/ml Hoeehst33342 then mounted on the slides to evaluate nuclear morphology using a microscope equipped with fluorescent illumination. [ Result] PMSG supplemented with the cumulus expansion was better. However, the percentages of MI-M 11 in PMSG groups had no significant differences (P 〈 0.05 ). GVBD of 100 IU/ml PMSG group was much lower than that in the control group. [ Conclusion] PMSG supplement in IVM medium could increase cumulus cell expansion, while the nuclear maturation rate remained low. Further studies were required to improve the nuclear development rate of canine oocytes.
出处
《安徽农业科学》
CAS
北大核心
2008年第35期15503-15504,15509,共3页
Journal of Anhui Agricultural Sciences