颗粒细胞双阶段共培养模式对性成熟前绵羊卵母细胞体外成熟的影响  

Effect of Biphasic Granulosa Cells Co-Culture on in vitro Maturation of Prepubertal Sheep Oocytes

作　　者：王海波 孙鹏 李玉杰 潘庆杰[1] 董焕声[1] WANG Haibo;SUN Peng;LI Yujie;PAN Qingjie;DONG Huansheng(College of Animal Science,Qingdao Agricultural University,Shandong Qingdao 2661091,China;Shandong Vocational Animal Science and Veterinary College,Shandong Weifang 261061,China)

机构地区：[1]青岛农业大学动物科技学院,山东青岛2661091 [2]山东畜牧兽医职业技术学院,山东潍坊261061

出　　处：《中国畜牧杂志》2021年第11期168-172,共5页Chinese Journal of Animal Science

基　　金：山东省动物种质资源保护与利用研究创新团队项目;山东省现代农业产业技术体系羊产业创新团队项目(SDAIT-10-03);山东省农业重大应用技术创新项目(6682217004)。

摘　　要：为改善性成熟前绵羊卵母细胞的体外成熟质量和发育能力,本研究通过构建颗粒细胞双阶段共培养模式来优化性成熟前绵羊卵母细胞体外成熟(IVM)体系,为利用颗粒细胞-卵母细胞双阶段共培养模式来改善卵母细胞体外发育能力提供依据。本实验以常规方法IVM 24 h作为对照,双阶段共培养组在以颗粒细胞为滋养层与卵母细胞共培养6 h后,继续用常规成熟培养基IVM至27 h。统计所有处理组IVM后的卵母细胞进行核成熟率、卵母细胞老化、线粒体分布情况和活性氧(ROS)水平。结果表明:颗粒细胞双阶段共培养组的卵母细胞核成熟率(58.15%)略高于对照组(P>0.05);线粒体呈皮质/核周分布的比例(56.25%)高于对照组(P<0.05);而卵母细胞的老化率(17.40%)、胞质内ROS水平(9.78%)均低于IVM 27 h组卵母细胞(P<0.05),与对照组无显著差异。综上所述,颗粒细胞双阶段共培养模式可以解除颗粒细胞对减速分裂的抑制作用,促进线粒体正确分布,抑制胞质内ROS水平升高,减缓因延长IVM时间造成的卵母细胞老化情况。In order to improve the quality and development ability of prepubertal sheep oocytes in vitro maturation,here we constructed the biphasic granulosa cells co-culture model to optimize the system of prepubertal sheep oocytes in vitro maturation(IVM),aiming to provide a basis for the use of this biphasic co-culture mode to improve the in vitro development of oocytes.The conventional method IVM 24 h was as a control,and in the biphasic co-culture group,the oocytes were co-cultured with granulosa cells feeder layer for 6 h,and then conventional maturation medium IVM 27 h was used continuously.The nuclear maturity rate,oocyte aging,mitochondrial distribution and reactive oxygen species(ROS)levels of oocytes in all groups after IVM were identified and counted.The results showed that the nuclear maturation of oocytes in biphasic granulosa cells co-cultured group was 58.15%slightly higher but not significantly different(P>0.05);The ratio of pericortical/perinuclear mitochondria distribution was 56.25%,which was significantly higher than that of the control group(P<0.05);the aging rate of oocytes was 17.40%,and the level of ROS in the cytoplasm was 9.78%,which were significantly lower than that of the oocytes in the IVM 27 h group(P<0.05)and were not significantly different from that of the control group(P>0.05).In summary,the biphasic granulosa cells co-culture mode can relieve the inhibitory effect of granulosa cells on decelerated division of oocytes,promote the correct distribution of mitochondria,inhibit the increase of ROS levels in the cytoplasm,and slow down the aging of oocytes caused by prolonged IVM time.

关 键 词：颗粒细胞 双阶段共培养 性成熟前绵羊 卵母细胞 

分 类 号：S826.3[农业科学—畜牧学]