中文摘要：

成熟卵母细胞的细胞质对供体细胞的重编程有作用,卵母细胞的成熟状态成为核移植过程的关键因素.为了研究卵母细胞体外成熟过程中卵母细胞生发泡破裂（germinal vesicle breakdown,GVBD）发生前后脱卵丘细胞对卵母细胞的影响,本研究采用猪（Sus scrofa）卵丘-卵母细胞复合体（cumulus-oocyte complexes,COCs）于体外培养,每隔5h取卵母细胞采样,置于地衣红染液中染色,根据卵母细胞染色体构型的变化确定卵母细胞的分期,然后在21、27和42 h的卵母细胞进行脱卵丘细胞处理,观察极体数.体外培养不同时间压片观察结果表明,直径2～6 mm卵泡卵母细胞在体外培养时,2～17 h大部分卵母细胞处于GV期（88.30％～52.38％）; 22 h有63.25％的卵母细胞处于生发泡破裂至前中期（germinal vesicalbreakdown～premetaphase Ⅰ,GVBD～pre-M Ⅰ）;27 h处于中期Ⅰ（metaphase Ⅰ,MⅠ）的卵母细胞最多（42.25％）; 32 h处于后期Ⅰ（anaphase Ⅰ,AⅠ）的卵母细胞比例最多（29.90％）; 37 h处于末期Ⅰ（telophase Ⅰ,TⅠ）的卵母细胞比例最多（40％）;42 h绝大部分卵母细胞达到了中期Ⅱ（metaphaseⅡ,MⅡ）（72.81％）.分别采取21、27和44 h的卵母细胞进行脱卵丘细胞处理,发现21h脱卵丘细胞后卵母细胞成熟率为25.56％,低于27 h处理卵的成熟率（34.33％）,42 h脱卵丘细胞的卵母细胞的成熟率最高达到84.33％.结果说明,体外卵母细胞成熟过程中,卵丘细胞对卵母细胞的成熟具有重要意义,为进一步探讨和分析猪卵母细胞体外成熟培养的方法,改良体外培养体系,提高猪卵母细胞体外培养成熟率,进而提高胚胎体外培养质量提高奠定基础.

英文摘要：

The mature oocyte cytoplasm have a role in the reprogramming of donor cells, maturation of oocytes become the key factors in the process of nuclear transplantation. In order to study the effect of the removal of cumulus cells around germinal vesicle breakdown（GVBD） on porcine（Sus scrofa） oocyte in vitro maturation, porcine cumulusoocyte complexes （COCs） were cultured in vitro and got samples per 5 h. Chromosomal morphological changes of porcine oocyte in vitro maturation were examined by aceto-orcein staining technique, and established the timetable of the oocytes chromosome developmental process. Then the cumulus cells were carried off from COCs in 21, 27 and 42 h, respectively. The effects of cumulus cells on oocyte maturation were investigated by counting the number of excreted polar body. From 2-17 h, most of the oocytes were in GV stage （88.30%-52.38% ）; 22 h, most of the oocytes were in germinal vesical breakdown-premetaphase I （GVBD~pre-M I ）stage（63.25%）; 27 h, most of the oocytes in metaphase I （M I ） stage（42.25%）; Until culturied for 32 h, the majority oocytes stayed in anaphase I （A I ）（29.90%）; 37 h, telophase I （T I ） oocytes were the most （40%）; And by 42 h, most of the oocytes reached metaphase 1/（M l/） stage （72.81%）. We carried off cumulus cells at 21, 27 and 44 h and set these oocytes as 3 groups. The result showed that the maturation rate of 21 h group was 25.56%, lower than that of 27 h group, which reached 34.33%, the 42 h group was the highest that reached 84.33%. These results illustrate the importance of cumulus cells on oocyte maturation in vitro oocyte maturation. This study is conducive to further exploring and analyzing the methods of porcine oocytes matured in vitro, improving the culture system for enhanced maturation rate of porcine oocytes in vitro, and lay the foundation for the quality of embryos cultured in vitro.