农业生物技术学报2012，Vol.20Issue(4)：411-419,9.DOI:10.3969/j.issn.1674-7968.2012.04.010
小鼠卵丘细胞对牛卵母细胞体外成熟和孤雌发育的影响
Effects of Mouse Cumulus Cells on Bovine Oocytes Maturation In vitro and Development after Parthenogenesis
摘要
Abstract
To research the effects of heterogeneous mouse cumulus cells (mCCs) on in vitro maturation of bovine denuded oocytes (DOs) and its development after parthenogenesis, bovine denuded germinal vesicle (GV) stage DOs were co-cultured in vitro with a mCCs monolayer in this study. The effect mechanism of cumulus cells on bovine DOs after in vitro maturation (IVM) and parthenogenesis were also discussed, and this could help to provide experimental bases for increasing maturation rate of bovine oocytes in vitro. Bovine oocytes were divided into four groups, namely COCs (cumulus-oocytes complexes) group, DOs+bCCs (bovine cumulus cells) group, DOs+mCCs group and DOs group. The extruded rate of oocytes in different groups of first polar body, glutathione (GSH) content and parthenogenetic development of M TI stage oocytes, relative expression levels of bone morphogenetic protein 15 (BMP1S) and growth differentiation factor 9 (GDF9) mRNA of oocytes were detected after in vitro mature for 22~24h. The results showed that (1) Nuclear maturation rate of DOs+mCCs group and DOs+bCCs group [(68.48±5.71)%, (71.00±4.27)%] were significantly lower than that of COCs group (84.70±3.41)% (P0.05), but significantly higher than that of DOs group (54.26±5.03)% (P0.05); After parthenogenetic activation the GSH content and blastocyst rate of M Ⅱ oocytes of DOs+mCCs group [ (2.48±0.24)pmol/oocyte, (44.19±4.03)%] were similar to those of DOs+bCCs group [(2.49±0.18)pmol/oocyte, (48.57±5.20)%] and DOs group [(2.40±0.17)pmol/oocyte, (43.47±7.34)%], but were all significantly lower than those of COCs group [(9.67+0.18)pmol/oocyte, (59.93±6.13)%)](P<0.05), and cleavage rate of different groups showed no significant difference [(98.85±4.32)%, (92.11+2.00)%, (95.40±4.11)%, (93.18+3.19)%] (P>0.05); (3) Relative expression levels of BMP15 and GDF9 mRNA of DOs+mCCs, DOs+bCCs and COCs group were all significantly higher than that of DOs group (P<0.05), but relative expression levels of GDF9 mRNA of DOs+ mCCs and COCs group were significantly higher than that of DOs+bCCs group (P<0.05). The result indicated that mCCs could improve nuclear maturation rate and BMP15, GDF9 mRNA relative expression levels of bovine DOs, but could not enhance cytoplasmic maturation of DOs and parthenogenetic embryo development of M Ⅱ oocytes. It's suggested that cumulus cells can promote bovine oocytes nuclear maturation in vitro through some paracrine factors, and the effects of these factors may have no species specificity.关键词
牛/小鼠/卵丘细胞/共培养/体外成熟/孤雌发育Key words
Bovine/ Mouse/ Cumulus cells/ Co-culture/ in vitro mature/ Parthenogenetic development引用本文复制引用
任晶晶,赵学明,秦彤,杜卫华,郝海生,刘岩,王栋,朱化彬..小鼠卵丘细胞对牛卵母细胞体外成熟和孤雌发育的影响[J].农业生物技术学报,2012,20(4):411-419,9.基金项目
国家奶牛产业技术体系建设专项资金(No.CARS-37)和国家外国专家局引智示范项目(No.Y20110326005) （No.CARS-37）
