中文摘要：

目的改良人羊膜上皮细胞的培养方法,检测生长特性,观察荧光染料4,-6-联脒-2-苯基吲哚（DAPI）标记人羊膜上皮细胞（HAECs）的效率。方法取足月剖宫产术后羊膜经胶原酶和胰蛋白酶消化获得上皮细胞。MTT法绘制细胞生长曲线、流式细胞仪检测细胞的生长周期,免疫组化对细胞角蛋白进行鉴定。并用DAPI标记,测1、2周荧光阳性率。结果培养的HAECs多角形,生长分为缓滞期、对数增长期和平台期,近80%处于细胞周期的静止期G0。细胞角蛋白keratin阳性表达。DAPI标记1周标记率为100%;2周细胞形态好,标记率不变,荧光强度减弱。结论该实验方法成功培养人羊膜上皮细胞。DAPI短期标记效果理想。

英文摘要：

Objective To improve the culture and DAPI labeling of human amniotic epithelial cells（hAECs） in vitro.Methods The hAECs isolated from amniotic membranes of uncomplicated elective caesarean section were digested by collagenase and trypsin.Cell viability and cell cycle was assessed with MTT and flow cytometry,respectively.And cytokeratin was detected with immunohistochemistry.DAPI staining was performed for assessment of fluorescence efficacy at 1w and 2w.Results Pylygonal hAECs was observed in stationary-phase,log-phase and plateau-phase with 80% stayed in G0 phase.Cytokeratin immunohistochemistry staining was positive.All hAECs were successfully labeled with DAPI at 1st and 2nd week with fluorescence intensity reduction at 2nd week.Conclusion Human AECs were successfully cultured with well DAPI labeled.