绵羊子宫内膜上皮细胞的纯化培养与鉴定  

Cultivation, Purification and Characterization of Ovine Endometrial Epithelial Cells

作　　者：马欣[1] 邢书涵[1] 刘云海[1] 郭勇[1] 倪和民[1] 

机构地区：[1]北京农学院动物科学技术学院,北京102206

出　　处：《中国农学通报》2015年第11期63-67,共5页Chinese Agricultural Science Bulletin

基　　金：2013年度北京市教委北京市属高等学校创新团队建设与教师职业发展计划项目"体细胞转基因克隆肉牛新品系培育与利用"(PXM2013_014207_000067);2013年度国家自然基金面上项目"牛体内外植入前胚胎IFNτ的差异表达及对子宫上皮细胞ISG15和Wnt7a表达的体外诱导"(31272526)

摘　　要：为了探索一种分离纯化绵羊子宫内膜上皮细胞的简便高效方法,采用先组织块培养后消化纯化和直接消化后过筛纯化2种方法对绵羊子宫内膜上皮细胞进行培养、纯化,最后用免疫荧光染色方法对上述方法得到的细胞进行角蛋白检测从而鉴定上皮细胞纯度。结果表明：先组织块培养后消化纯化法可以得到纯度95%,且形态均一、活性良好的绵羊子宫内膜上皮细胞;先消化后过筛纯化法得到的子宫内膜上皮细胞形态良好但是纯度仅为70%,与前一方法相比有待提高。因此,先组织块培养后消化纯化法是获得绵羊子宫内膜上皮细胞的一种简单易行且高效的方法,该方法得到的细胞可以在体外传至3～4代。The objective of this study was to investigate a simple and efficient way to obtain purer ovineendometrial epithelial cells cultured in vitro. Two methods, which were trypsin digestion following tissueculturing and filtration method following collagenase digestion, were applied to cultivate and purify ovineendometrial epithelial cells. Then, cytokeratin was detected with immunocytochemical staining to evaluate thepurity of ovine endometrial epithelial cells collected by the two different ways. The results showed that highpurity（95%） of epithelial cells which exhibited homogeneous morphology and good cell activity could beobtained using trypsin digestion following tissue culturing. However, purity of epithelial cells（about 70%） waslower using filtration method following collagenase digestion even though the morphology of cells was similar tothat of former one. Therefore, it could be concluded that trypsin digestion following tissue culturing was asimple and efficient way to purify ovine endometrial epithelial cells which could be passaged for at least 3-4generations when cultured in vitro.

关 键 词：绵羊 子宫内膜上皮细胞 纯化 

分 类 号：S814.8[农业科学—畜牧学]