人尿促性腺激素、雌二醇及表皮生长因子对牛卵母细胞体外成熟质量的影响

Effect of Human Menopausal Gonadotropin,17β-estradiol and Epidermal Growth Factor on the Quality of in vitro Maturation Bovine Oocytes

试验旨在研究不同激素配比及表皮生长因子(EGF)浓度对牛卵母细胞体外成熟及卵母细胞质量的影响。将随机分组的卵丘-卵母细胞复合体于添加FSH+LH、HMG、FSH+LH+E2、HMG+E24种不同激素组合配比的成熟基础液中培养,对比其体外成熟率,比较了EGF对牛卵母细胞体外成熟率和孤雌胚胎体外发育的影响,并采用TUNEL法检测添加不同浓度EGF的牛孤雌激活囊胚细胞凋亡情况。结果表明,添加HMG的成熟试验结果稳定,E2对牛卵母细胞成熟有一定的促进作用,HMG+E2联合使用可以得到高效稳定的成熟结果;在此基础上,在成熟液中添加30ng/mL EGF对牛卵母细胞的成熟质量、胚胎发育及降低胚胎细胞凋亡都有明显的促进作用。因此,在体外成熟培养液中添加0.075IU/mL HMG、1μg/mL E2和30ng/mL EGF对牛卵母细胞的成熟和质量较为有益。

For optimizing in vitro maturation system of bovine oocytes,we firstly examined the influence of four different hormonal regimes（FSH＋LH,HMG,FSH＋LH＋E2and HMG＋E2）on oocyte maturation rates.Then we studied the effects of epidermal growth factor（EGF）in the above defined medium on bovine oocyte maturation,in vitro development and quality of parthenogenetic embryos.The cell apoptotic index of parthenogenetic blastocysts was detected by TUNEL.No significant difference was observed in maturation rates in four groups supplemented with different hormones.However,human menopausal gonadotropin（HMG）provided steady maturation results in replicates.Maturation of oocytes was promoted by supplementation with17β-estradiol（E2）.Combination of HMG and E2 gave rise to steady and efficient mature results.The presence of EGF at 30ng/mL concentration significantly increased maturation rate and blastocyst rate and reduced apoptotic cells in parthenogenetic blastocysts.Therefore,the optimal oocyte maturation solution could be supplemented with 0.075IU/mL HMG,1μg/mL E2 and 30ng/mL EGF.