小鼠腔前卵泡无血清体外培养的研究

Study of serum free culture in vitro for mouse preantral follicles and oocytes

目的:探索添加物维生素C和促性腺激素对小鼠腔前卵泡卵母细胞体外生长发育的影响及适合小鼠腔前卵泡体外生长发育的培养体系.方法:通过改良的TCM199无血清培养系统,在96孔细胞培养板中,每孔放入1个腔前卵泡等条件下体外培养腔前卵泡卵母细胞.结果:随着培养时间的延长,各组卵泡和卵母细胞的直径都不同程度的有所增长,其中FSH400+LH200组和VC50+FSH400+LH200影响均大于VC50和对照组,存在显著性差异(P<0.05),但二者之间无显著性差异.FSH400+LH200质量浓度组卵泡存活率、发生GVBD率和第一极体排出率均最高,分别为68.6%、17.6%、9.8%.但VC50+FSH400+LH 200组卵泡成腔率最高,为48%.结论:在改良的TCM199无血清培养系统中,联合添加促性腺激素和抗氧化剂后,卵泡成腔率大有提高,卵泡存活率、发生GVBD率和第一极体排出率仅次于单独添加促性腺激素组.

Aim:Investigating the influence of antioxidant vitamin C and gonadotropin on the growth and development of mouse preantral follicle oocytes in vitro,in order to explore a suitable growth culture system for mouse preantral follicles in vitro.Methods:Using the modified TCM199 serum-free culture system,one preantral follicle was placed in each well of a 96-well culture plate for culture of the preantral follicular oocytes in vitro.Results:With culture time,the diameter of follicle and oocyte increased in varying degrees.Groups FSH400 + LH200 and VC50 + FSH400 + LH200 were found to have greater effects than those of VC50 and the control groups,with significant difference(P < 0.05),but no significant difference was found between the two groups.The FSH400 + LH200 group had highest rates of follicle survival,GVBD and the first polar body,i.e.68.6%,17.6%,9.8%,respectively.But follicular cavity of the VC50 +FSH400 +LH 200 group was found to be the highest,i.e.48%.Conclusion:In the modified TCM199 serum-free culture system,follicular cavity rate increased greatly after combined addition of gonadotropin and antioxidants,but the follicle survival rate,GVBD rate and the first polar body rate were lower than those of the group with addition of only gonadotropin.