兔卵母细胞体外成熟培养影响因素的研究  

Study on the Influencing Sectors in Vitro Maturation of Rabbit Oocytes

作　　者：王辉田[1] 罗光彬[1] 王忠彬[1] 孙超[1] 陈思[1] 

机构地区：[1]沈阳农业大学畜牧兽医学院,沈阳110866

出　　处：《中国养兔》2014年第2期7-12,共6页Chinese Journal of Rabbit Farming

摘　　要：试验旨在探讨卵丘细胞包裹层数、表皮生长因子(epidermal growth factor,EGF)和谷胱甘肽(glutathione,GSH)对兔卵母细胞体外成熟培养的影响。分别采用切割法和针刺挤压法取母兔卵巢表面的卵丘卵母细胞复合体(cumulus oocyte complexes,COCS),将获取的COCS在基础培养液中添加不同浓度表皮生长因子(0ng/mL、10ng/mL、20ng/mL、30ng/mL)及不同浓度的谷胱甘肽(0μmol/mL、0.5μmol/mL、1.0μmol/mL、1.5μmol/mL、2.0μmol/mL)进行体外成熟培养并观察第一极体排出率。结果表明:①手术刀切割法和针刺挤压法平均回收卵母细胞率分别为(22.83、39.29)枚,针刺挤压法显著高于手术刀切割法(P【0.05)。②3层以上卵丘细胞包裹的卵母细胞体外成熟率为81.27%,显著高于1~3层卵丘细胞包裹的体外成熟率63.75%和裸卵细胞体外成熟率23.45%(P【0.05)。③添加20ng/mL的EGF对兔卵母细胞体外成熟培养后成熟率为82.14%,效果最好,且显著高于对照组10ng/mL和30ng/mL EGF组(P【0.05)。④添加不同浓度的谷胱甘肽可以促进和提高兔卵母细胞体外成熟率,其中添加1.0μmol/mL的谷胱甘肽时,卵母细胞成熟率为71.44%,与添加1.5μmol/mL组的相比差异不显著(P】0.05),但显著高于其他组(P【0.05)。结论为添加20ng/mL的EGF和1.0μmol/mL谷胱甘肽,对用针刺挤压法获得3层以上卵丘细胞包裹的兔卵母细胞体外培养效果最佳。To study the effect of cumulus cell,epidermal growth factor(EGF)and glutathione(GSH)on in vitro maturation culture of rabbit oocytes. Cumulus oocyte complexes(COCS)were collected from the surface of ovarian by the method of slicing using scalpel or by stab-extruding with a needle attached to a syringe. In vitro cumulus oocyte complexes were cultured maturation in basic culture medium supplemented with different concentrations of EGF(0 ng/ mL,10 ng/ mL,20 ng/ mL,30 ng/ mL)and different concentrations of GSH(0 μmol/ mL,0.5 μmol/ mL,1.0 μmol/ mL,1.5 μmol/ mL,2.0 μmol/ mL).The proportion of oocytes in the first polar body expelled was observed. The results showed that:①The average recovery rates were 22.83 and 39.25 oocytes per rabbit by the method of slicing and stab-extruding respectively,and the average number recovered rate of per rabbit by tab-extruding was significantly higher than those by slicing(P<0.05). ②The oocytes maturation rate of cumulus cells covered with more than 3 layers was 81.27%,andwas significantly higher than that of cumulus cells covered with 1~3 layer(63.75%),and the cumulus maturation without layer(23.45%)(P<0.05). ③The maturation medium supplemented with the EGF group was significantly higher than that of control group(P<0.05).The best results was obtained from the supplemented with 20 ng/mL EGF in the maturation medium, and the vitro maturation rate of rabbit oocytes was 82.14.There was significant difference between 10 ng/mL and 30 ng/mL EGF group(P<0.05). ④Supplement different concentrations of GSH can promote and increase the maturation rate of rabbits oocytes in vitro. The oocytes maturation rate was 71.44% by 1.0 μmol/ mL GSH supplemented,and had no significantly different by 1.5 μmol/mL GSH(P>0.05),but it was significantly higher than other groups(P<0.05). These results indicated that with the maturation medium supplemented with 20 ng/mL EGF and 1.0 μmol/ mL,and by using the method of stab-extruding with a needle attached to a syringe, it had better effect on more th

关 键 词：兔 卵母细胞 体外培养 影响因素 

分 类 号：Q813[生物学—生物工程]