人羊膜上皮细胞的分离、培养及鉴定

目的建立人羊膜上皮细胞分离、培养及鉴定方法。方法从足月分娩人胎盘剥离羊膜，采用胰蛋白酶消化法分离人羊膜上皮细胞（hAECs），采用免疫细胞化学、免疫荧光染色及流式细胞术分析其表型特征，使用含表皮生长因子LG—DMEM培养基培养。结果采用低速旋转胰蛋白酶消化法从羊膜分离的hAECs数为（6．13±1．42）×10^7，份（n=12），所分离的hAECs明显表达上皮细胞标志物角蛋白CK19，不表达间充质细胞标志物波形蛋白，不同程度地表达CD29、CD44和CD71。在表皮生长因子存在的条件下，hAECs生长增殖较快，培养至第6天细胞总数可增殖171倍。结论建立了人羊膜上皮细胞分离、培养及鉴定方法，hAECs具有上皮细胞的特异标志和间充质干细胞的某些表型特征。

Isolation, culture and identification of the amniotic epithelial cells derived from human amnion

Objective To investigate the isolation and culture condition of human amniotie epithelial ceils （hAECs） and its phenotypic identification. Methods The hAECs were isolated from human amnion digested with trypsin.The phenotype of hAECs was identified by Immunocytochemistry, immunofluorescence and flow cytometry. The cells were cultured in LG- DMEM medium containing epidermal growth factor. Results A term amnion may yield uo to （6.13±1.42）×10^7 hAECs by low velocity-trypsin digestion .Cytokeratin 19 was expressed in the amniotic epithelial ceils derived fl＇om human amnion, while vementin was negative , and CD29、CD44 and CD71 different extenfly were expressed . The total cell number hAECs was expanded by 167-fold after culture for 6 days.Conclusions A method for isolation, culture and identification of hAECs derived from human amnion is successfully established. The hAECs have specific marker of epithelial cells and some phenotypic properties of mesenchymal stem cells.