原代人子宫内膜细胞分离及培养鉴定

目的：探究原代人子宫内膜细胞分离及培养的简易方法，并对其纯度及生物活性进行鉴定。方法采用胶原酶消化人子宫内膜组织，经2次筛网过滤、离心及贴壁纯化技术，分离、纯化培养人子宫内膜间质细胞（ESC ）和腺上皮细胞（EEC ），用细胞角蛋白（cytokeratin）和波形蛋白（vimentin）免疫细胞化学染色法和免疫荧光方法对所分离的细胞进行鉴定。结果 ESC呈平行生长，细胞呈梭形或多角形，波形蛋白抗体免疫细胞化学染色呈阳性，纯度达95％以上。EEC呈漩涡状生长，细胞呈多角形或蝌蚪形，细胞角蛋白抗体免疫细胞化学染色呈阳性，纯度可达90％。结论应用胶原酶消化法及二次筛网过滤法成功分离并培养数量、活力和纯度高的子宫内膜细胞，可操作性强，可供具备基本细胞培养条件的实验室进行推广。

The method of isolation,culture and identification of primary human endometrial cells

Objective To study the isolation method and the culture method of primary human endometrial cells and to iden‐tify the purity and biological activity .Methods We digested human endometrial tissue by collagenase at first ,and then to separate , purify and culture the human glandular stromal cells(ESC) and endometrial epithelial cells(EEC)by differential centrifugation ,dab‐ble screen filtration and adhesion purification technology .Ultimately ,we identified the isolated cells with cytokeratin and vimentin immunocytochemical staining and immunofluorescence method .Results Stromal cells showed a parallel growth .The cells were spindle or polygonal ,and the vimentin antibody showed a positive immunohistochemical staining ,the purity was more than 95% .At the same time ,glandular epithelial cells grew in whorls .The cells were polygonal or tadpole shaped ,and the cytokeratin antibody immunohistochemical staining ,the purity were up to 90% .Conclusion The successful isolation and culture of high quantity ,viabili‐ty and purity by collagenase digestion and dabble screen filter method of endometriall cells make a strong operability .The laboratory which has the basic cell culture conditions can develop the experiment .