培养时间和表皮生长因子对兔卵母细胞体外成熟的影响

Effect of Culture Time and Epidermal Growth Factor on the in vitro Maturation of Rabbit Oocytes

本试验旨在探讨培养时间和表皮生长因子(EGF)对兔卵母细胞体外成熟及受精的影响。分别用切割法和针刺挤压法取经超排后母兔卵巢表面的卵母细胞颗粒细胞复合体(COCs),将COCs分别在不同时间(24、30、36及40h)和在基础培养液中添加不同浓度表皮生长因子(0、50、100ng/mL)进行体外成熟培养并观察其第一极体排出率。结果表明:①针刺挤压法获得COCs数明显高于切割法(P<0.05);②体外成熟培养时间30(36)h的第一极体排出率高于24(40)h(P<0.05);③添加50和100ng/mLEGF组卵母细胞第一极体排出率明显高于对照组(P<0.05)。因此,①兔卵巢卵母细胞的获得方法以针刺挤压法较好;②兔卵母细胞成熟时间以30～36h为宜;③EGF对兔卵母细胞体外成熟有促进作用,在基础培养液中添加100ng/mLEGF为宜。

To study effect of culture time and epidermal growth factor on the in vitro maturation of rabbit oocytes. After superovulating female rabbit,we killed it and took out it’s ovarians,washed them and obtained COCs on the surface of ovarian by method of cutting with blade or by stab-extruding with niddle,observed the emission rate of the first polar body at different times （24,30,36 and 40 hours） and added in the basic culture medium with different concentrations （0,50,100 ng/mL） of epidermal growth factor （EGF） in vitro maturation. The results showed that： ①Method of stab-extruding with niddle to obtain the number of COCs was bitter than cutting with blade （P〈0.05）; ②the first polar body emission rate at in vitro maturation time of 30 （36） hours of was higher than 24 （40） hours （P〈0.05）; ③the first polar body emission rate in adding 50 ng/mL and 100 ng / mL EGF group was significantly higher （P〈0.05）. Therefore,①Method of stab-extruding with niddle was better than method of cutting with blade in obtaining COCs on the surface of the rabbit ovarian;②On in vitro maturation of rabbit oocyte,time of 30 to 36 hours was appropriate;③EGF could significantly improve oocytes maturation,it was suitable to add 100 ng/mL EGF in the basic culture medium.