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中国农学通报

中国农学通报 ›› 2015, Vol. 31 ›› Issue (11): 63-67.doi: 10.11924/j.issn.1000-6850.casb14120034

所属专题： 生物技术； 畜牧兽医

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

绵羊子宫内膜上皮细胞的纯化培养与鉴定

马欣,邢书涵,刘云海,郭勇,倪和民   

  1. 北京农学院动物科学技术学院,北京农学院动物科学技术学院,北京农学院动物科学技术学院,北京农学院动物科学技术学院,北京农学院动物科学技术学院
  • 收稿日期:2014-12-03 修回日期:2015-03-30 接受日期:2015-03-25 出版日期:2015-05-06 发布日期:2015-05-06
  • 通讯作者: 倪和民
  • 基金资助:
    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)。

Cultivation,Purification and Characterization of Ovine Endometrial Epithelial Cells

倪和民   

  • Received:2014-12-03 Revised:2015-03-30 Accepted:2015-03-25 Online:2015-05-06 Published:2015-05-06

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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,本试验采用先组织块培养后消化纯化和直接消化后过筛纯化两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。

关键词: 硫磺粉, 硫磺粉

Abstract: The objective of this study was to investigate a simple but efficient way to obtain purer ovine endometrial epithelial cells cultured in vitro. In this experiment, two methods were applied as following: one is tissue culturing following trypsin digestion; the another one is collagenase digestion by filtration. Finally, cytokeratin 18 was detected with immunocytochemical staning to evaluate the purity of ovine endometrial epithelial cells collected by these two different ways. The result showed that by method of tissue culturing following trypsin digestion, we could obtain high purity epithelial cells (>95%), which exhibited homogeneous cobblestone morphology and good cell activity. Compared with this method, the collagenase-digesting way by filtration could get lower purity epithelial cells (about 70%), even though the morphology of cells was similar to that of formal one. Collectively, it can be concluded that tissue culturing with trypsin digestion is a simple and efficient way for purifying ovine endometrial epithelial cells, which can be passaged for at least 3-4 generations while they are cultured in vitro.