绵羊子宫内膜上皮细胞的纯化培养与鉴定

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。

Cultivation,Purification and Characterization of Ovine Endometrial Epithelial Cells

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.