人子宫内膜细胞体外培养模型的建立

Establishment of Human Endometrial Cell Culture Model in Vitro

【目的】构建人子宫内膜细胞体外培养模型。【方法】收集子宫内膜组织，采用酶消化、筛网滤过、梯度离心的方法获得腺上皮与基质细胞混合细胞进行体外原代培养，采用光学显微镜下观察体外培养的细胞形态，采用波形蛋白和角蛋白单克隆抗体进行免疫组化法鉴定子宫内膜基质细胞和腺上皮细胞。【结果】倒置显微镜下发现子宫内膜细胞中腺上皮细胞和基质细胞呈混合生长，其中基质细胞占大部分，为72．0％；腺上皮细胞占小部分，为24．5％，基质细胞和腺上皮细胞免疫组化染色后呈阳性，细胞纯度分别为24．5％和70．5％，两种细胞加起来的纯度为95％。【结论】该方法操作简便，效率高，可建立一个稳定实用的人子宫内膜细胞体外培养体系。

[Objective] To explore the methods of isolating and identifying human endometrial cells for es‐tablishing a stable in intro culture system of endometrial cells .[Methods] The methods of collagenase diges‐tion ,sieve filtration and gradient centrifugation were employed for isolating epithelial and stromal cells for pri‐mary culturing .Cellular morphology was observed microscopically .Endometrial stromal cells （ESCs） and en‐dometrial epithelial cells （EECs） were identified by immunohistochemistry with cytokeratin and vimentin mon‐oclonal antibodies respectively .[Results] Endometrial cells could be isolated completely .Both EEC and ESC existed in culture and stained positively by immunohistochemistry .The purity of both cells were 24 .5% and 70 .5% respectively with a total purity of 95% .[Conclusion] A stable in vitro culture system of human endo‐metrial cells has been established for elucidating the physiologic and pathologic mechanisms of human endome‐trium and drug actions .