中文摘要：

目的：探讨人羊膜上皮细胞（human amniotic epithelial cells,HAECs）诱导分化为角膜上皮细胞的可塑性及其重建角膜上皮的可行性。方法：取足月产人羊膜,通过胶原酶、胰蛋白酶和EDTA消化获得HAECs,将细胞在体外培养扩增、传代,将2～3代HAECs接种于去上皮的兔角膜基质上培养,待HAECs融合形成单层后,置于插入式培养皿中进行气液界面培养14 d。对角膜基质上培养的HAECs用光镜、扫描电镜和透射电镜进行形态学及超微结构观察,以及细胞角蛋白（CK）3/12的免疫组织化学检测。结果：HAECs可在角膜基质上生长,培养1～2 d可形成单层,气液界面培养14 d可形成4～5层细胞的复层上皮,扫描电镜观察细胞表面有丰富的微绒毛,透射电镜下可见上皮细胞之间有桥粒连接,免疫组化检测复层细胞表达CK3/12,所形成的复层结构与正常角膜上皮相似。结论：HAECs有可能作为种子细胞用于组织工程角膜上皮重建。

英文摘要：

AIM： To investigate the plasticity of human amniotic epithelial cells induced to differentiate into corneal epithelial cells and the feasibility of corneal epithelium reconstruction.METHODS： A piece of amniotic membrane taken from an uncomplicated elective caesarean section was digested by collagenase,trypsin and EDTA.The amniotic epithelial cells were primarily cultured and passaged in Dulbecco＇s modified Eagle＇s medium containing 10% fetal bovine serum.The HAECs of the 2nd or 3rd generation were continuously cultured on fresh corneal stroma.One or two days later,the human amniotic epithelial cells formed a monolayer,and then the air-lifting cultivation was carried out in an insert culture dish.The cultured cells on corneal stroma were investigated morphologically by hematoxylin-eosin staining,and its ultramicrostructure was studied by scanning electron microscopy（SEM） and transmission electron microscopy（TEM）.Meanwhile,the expression of cytokeratin（CK）3/12 was detected by immunohistochemistry.RESULTS： Human amniotic epithelial cells were successfully reproduced on corneal stroma in vitro.The monolayer was formed after cultured for 1 or 2 days,and stratification with 4 to 5 layers of epithelial cells was observed 14 days after air-lifting cultivation.A wealth of microvilli on the surface of HAECs were observed by SEM,while desmosomes among HAECs were found by TEM.The stra-tified cells expressed CK3/12.The composed tissue was very similar to normal corneal epithelium.CONCLUSION： Human amniotic epithelial cells can be used as seed cells to reconstruct the corneal epithelium by the technique of tissue engineering.