输卵管上皮细胞共培养对小鼠早期胚胎发育阻滞的影响  

Effect of Oviduct Epithelial Cells Co-culture on the Developmental Arrest of Early Embryos in Mouse

作　　者：庄利利[1] 吴俊波[1] 胡德宝[1] 蔡丽娟[1] 张也[1] 李钟淑[1] 方南洙[1] 

机构地区：[1]延边大学农学院动物科学系,延吉133002

出　　处：《畜牧兽医学报》2013年第7期1160-1166,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：延边大学211工程第三期重点学科建设项目(2009-002);动物高效快繁及胚胎工程高新技术研究

摘　　要：研究输卵管上皮细胞共培养对小鼠早期胚胎发育的影响,探索输卵管上皮细胞共培养克服胚胎发育阻滞的机理。建立纯化的小鼠输卵管上皮细胞系,并与合子进行共培养,用DCHFDA测定胚胎内部的H2O2含量,以MTT法测定输卵管上皮细胞活力,用RT-PCR检测CAT及GPX的转录状况。输卵管上皮细胞共培养能够显著提高2-细胞早期、4-细胞期胚胎发育率(P<0.05),囊胚率也有所提高(P>0.05)。输卵管上皮细胞共培养能够显著降低2-细胞早期胚胎内部的H2O2含量(P<0.05)。小鼠阻滞胚胎中有H2O2蓄积,输卵管上皮细胞共培养能够降低Ⅰ和Ⅱ型阻滞胚胎的比例(P>0.05),显著地提高Ⅲ型阻滞胚胎的比例(P<0.05)。当胚胎发育到2-细胞早期时共培养细胞活力上升(P>0.05),抗氧化基因CAT及GPX转录量增加(P>0.05)。以上结果表明,输卵管上皮细胞共培养通过细胞-细胞间的相互作用降低胚胎细胞内部的活性氧,克服胚胎发育阻滞,促进胚胎发育,提高胚胎质量。This experiment was conducted to study the effect of oviduct epithelial cells co-culture on the development of early embryos, and explored the mechanisms of oviduct epithelial cells co- culture on overcoming the developmental arrest in mouse. Mouse oviduct epithelial cell line was purified and used to co-culture the zygotes, H2O2 contents in embryos were measured by DCHF- DA, MTT was used to measure the activity of epithelial cells, RT-PCR was applied to test the transcriptions of CAT and GPX genes. Oviduct epithelial cells co-culture can significantly im- prove the early 2-cell and 4-cell rates （P〈0.05）, the blastocyst rate was also increased （P〈 0.05）. Oviduct epithelial cells co-culture can decrease the H202 content in early 2-cell embryos （P〈0.05）. Also there was a deposit of H2O2 in the arrested embryos, the rates of type I and [[ arrested embryos were decreased（P〈0.05） ,type m was increased（P〈0.05）by oviduct epithelial cells co-culture. On the other side, the cellular activity was improved （P〈0.05） when embryos developed to early 2-cell phase, at the same time, transcriptions of CAT and GPX were increased （P〈0.05）. Those results indicated that oviduct epithelial cell co-culture can improve the devel- opment of embryos by cell-cell contact to decrease the reactive oxygen species content in embry- os, overcome developmental arrest and promote embryo development.

关 键 词：输卵管上皮细胞 共培养 早期胚胎 阻滞 活性氧 

分 类 号：S814.8[农业科学—畜牧学]