不同培养条件对绵羊卵母细胞体外成熟的影响  

Effects of the different conditions on in vitro maturation culture of ovary oocytes in sheep

作　　者：徐燕霞[1] 曹会萍[1] 马呈瑞[2] 阚威[2] 李展[2] 张小宇[2] 胡少辉[2] 张连梅[1] 常卫华[2] 张勇[2] 

机构地区：[1]甘肃农业大学生命科学技术学院,甘肃兰州730070 [2]甘肃农业大学动物医学院,甘肃兰州730070

出　　处：《甘肃农业大学学报》2014年第2期35-40,共6页Journal of Gansu Agricultural University

摘　　要：通过使用不同的卵母细胞采集方法、不同的培养液、培养时间和细胞因子对绵羊卵母细胞的体外成熟培养的影响进行了研究.结果表明：切卵法和抽吸法对卵母细胞成熟的影响前者优于后者,差异极显著（P〈0.01）；培养液中添加FSH、LH 及FBS对绵羊卵母细胞的影响显著高于添加发情羊血清的且差异极显著（P〈0.01）；绵羊的卵母细胞培养20 h的成熟率极显著低于培养22 h和24 h的成熟率（P〈0.01）,培养22 h和24 h的成熟率差异不显著（P〉0.05）,培养24 h和培养26 h的成熟率差异极显著（P〈0.01）,培养26 h卵母细胞老化对随后的体外受精不利；成熟液（培养液＋FBS、LH、FSH、E2）中分别加入骨形态发生蛋白15（bone morphogenetie protein,BMP15）和成纤维细胞生长因子8 b（fibroblast growth factor 8b,FGF8b）2种细胞因子,成熟液中添加 BMP15组与添加FGF8b组卵母细胞的成熟率差异极显著（P〈0.01）,成熟液中添加 BMP15和对照组卵母细胞成熟率的差异极显著（P〈0.01）,加入 FGF8b组与对照组卵母细胞成熟率差异不显著（P〉0.05）.利用切卵法在培养液中加入 FSH、LHE2和细胞因子BMP15,培养22~24 h可以大大提高绵羊卵母细胞体外培养的成熟率.The influences of the different methods for harvesting the occytes,different culture matura-tion medias and various culture time on IVM of sheep oocytes were studied in this paper.The results showed that the effects of slicing on oocyte maturation were superior to the aspiration,and there were most significant differences between them （P〈0.01）.The effects of FSH,LH,FBS were better than those of ESS （P〈0.01）.It was higher in group which the oocytes were cultured for 22 and 24 hours compared with group that cultured for 20 hours （P〈0.05）,but there were no differences between 22 hours and 24 hours （P〉0.05）.There were most siginificant differences between 24 h and 26 h （P〈0.01）.It was harmful that the culture time of occyte was 26 h for the subsequential IVF.BMP15 （bone morphogenetie protein）and FGF8b（fibroblast growth factor 8b）were adder to the matural media.The mutural solution was regarded as the control group.The group of adding BMP15 had higher rate of IVM than the group of adding FGF8b＆amp;nbsp;（P〈0.01）,and the group of adding FGF8b and the control group had no difference.There were most sig-nificant differences between the group of adding BMP15 and the control group （P〈0.01）.Using the meth-od of slicing and adding FSH,LH,E2,BMP15 in the culture solution,the rate of occyte maturation of IVM of the sheep was very high after culturing for 22-24h.

关 键 词：绵羊 卵母细胞 体外成熟 BMP15 

分 类 号：S826[农业科学—畜牧学]