家兔原始卵泡卵母细胞体外发育的培养体系研究  

In Vitro Culture and Development of Rabbit Primordial Follicular Oocytes

作　　者：翟向玮[1] 孙理兰[1] 张攀[1] 潘庆杰[1] 沈伟[1] 

机构地区：[1]青岛农业大学动物科技学院动物生殖发育与基因工程研究所,山东青岛266109

出　　处：《青岛农业大学学报（自然科学版）》2009年第4期265-270,275,共7页Journal of Qingdao Agricultural University(Natural Science)

基　　金：国家863计划重点项目(2008AA101003);山东省自然科学基金(Z2007D01);山东省泰山学者岗位项目;青岛农业大学高层次人才启动基金重点项目(630615)

摘　　要：本研究根据家兔卵巢的发育特征初步探讨了家兔卵巢卵母细胞体外发育的培养体系。采集新生或30日龄家兔卵巢在不同培养液中培养,并分析家兔卵母细胞的体外发生与成熟情况与体内正常发育的兔卵巢卵母细胞之间的差异,来筛选和优化家兔卵巢卵母细胞的体外培养体系。本试验从高糖DMEM、FSH浓度、BSA/血清等3个方面来筛选较好的培养条件,结果表明:①不添加高糖DMEM的M199培养液与添加的相比,卵巢上卵母细胞数量要多,但随着培养时间的延长,组织块上卵母细胞的形态会变得不清楚;②家兔卵巢组织在FSH浓度为50和100m IU/m l、添加血清的DMEM/F12培养液中生长较好,体外培养24d后,卵母细胞数量多,直径增加到65μm;③与添加BSA的培养液相比,添加血清的培养液中的卵巢卵母细胞生长状态较好,体外培养28d后,卵母细胞数量多,并且直径可达65μm。In vitro culture of rabbit primordial follicular oocytes was investigated in this study. To select a optimal in vitro culture system of rabbit primordial follicle and oocytes, ovaries of newborn or day - 30 rabbits were cultured in different base - mediums plus FSH with distinct densities, and the ectogenesis and mature of oocytes were determined comparing with the same age in vivo. The better cuhure condition was selected from three aspects in our experiment, for example density of FSH,high glucose DMEM,BSA/FCS. The results show that： （1） there are more oocytes within the ovary tissues cultured in M199 without high glucose DMEM than that with high glucose DMEM, but then the shape of oocytes are more and more unclear in both two teams. （2） Ovaries cultured in DMEM/F12 and FCS plus 50 or 100 mIU/ml FSH grow were the best in all experimental groups, and with more and bigger oocytes even reaches 65 txm after 26 days in vitro culture. （3） The growth of ovaries cultured in the medium plus FCS was better than that of ovaries cultured in the medium plus BSA, and with more and bigger oocytes even reaches 65 μm in the former after 26 days in vitro culture.

关 键 词：家兔 卵巢 原始卵泡 卯母细胞 体外培养 

分 类 号：S814.4[农业科学—畜牧学]