摘要
为探明小鼠腔前卵泡体外发育及体外排卵的特征和规律性 ,为利用卵巢资源奠定基础 ,采用机械方法分离未成熟小鼠腔前卵泡进行体外培养 .以 α- MEM加亚硒酸钠、维生素 C,L-谷氨酰胺、丙酮酸钠为基础培养液 ,分组添加 10 %的 2种不同来源的血清和 1μg/ m L FSH.在多卵泡培养体系中 ,培养 9～ 10 d,每天观察其体外发育情况 ,测定卵泡和卵母细胞大小 .培养结束时 ,用 h CG进行体外诱导排卵 .结果表明 :体外培养过程中 ,腔前卵泡贴附培养皿底 ,相互聚集粘附形成团状结构 ,在细胞铺层微滴培养中 ,卵泡呈单个生长 ,并发育形成卵泡腔或腔样结构 ;经9d体外培养 ,小鼠腔前卵泡达到排卵前大小 (>40 0 μm) ,卵母细胞从开始时的 5 3.2 μm生长到 6 8.0～ 70 .8μm,总存活率为 6 1.0 % ;卵母细胞的大小及存活率在不同的血清添加组间差异不显著 ;用 h CG可诱导体外排卵 ,发生卵泡破裂 ,卵母细胞排出 ,卵丘细胞扩展 ,卵母细胞生化泡破裂 ,放出第一极体 .总排卵率达 5 9.5 % .因此 ,采用多卵泡培养体系 ,小鼠腔前卵泡能够发育至成熟排卵 .
Mechanically isolated early preantral mouse follicles were cultured in vitro to study their development and ovulation. Intact follicles were cultured in 100～150 microliters droplets α-MEM supplemented with sodium selenile,ascorbic acid,L-Glutamine,Sodium pyruate,sera derived from different animals(10%) and FSH(1 μg/mL) under paraffin oil at 37 ℃ and 100% humidity.During the culture period,the diameters of the follicles and oocytes were measured every day,and the development characterizations of the follicles were observed every day.At the end of the culture period,follicles were induced to ovulate with hCG.In this culture system the follicles were allowed to attach to the bottom of the dishes,in a cluster pattern;the follicles with an intact theca cell layer grew to form antral cavity, or the granulose cell outgrew to develop the follicles with antral-like cavity;after 9 days culture period, the preovutory stage was reached,and the oocyte diameter increased from 53.2 μm to 68.0～70.8 μm. A total survival rate of oocyte was 61.0%.There was no significant difference in the oocytes' growth and survival among sera derived from different animals (p>0.05).At the end of the culture period,follicles were ovulated in vitro after hCG addition, the ovulated oocytes exhibited cumulus expansion, germinal vesicle breakdown, and the first-popular body extrusion. The total ovulation rate was 59.5%.In summary, under the present culture conditions, the preantral follicles isolated from immature mice can mature and ovulate in vitro.
出处
《湖南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2001年第4期312-316,共5页
Journal of Hunan Agricultural University(Natural Sciences)
基金
国家教委优秀青年教师基金资助项目[教人司 (1994) 35 7号 ]
关键词
小鼠
腔前卵泡
体外发育
体外排卵
mouse
preantral follicle
in vitro development
in vitro ovulation
