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目的 观察皮肤干细胞在角膜基质上的分化发育情况 ,探讨皮肤干细胞重建角膜上皮的可能性。方法 将体外分离培养的 1～ 4代人和兔皮肤干细胞种植在兔角膜基质上 ,在体外和体内观察皮肤干细胞的分化发育情况。实验标本作HE染色及用人上皮细胞角蛋白K19单克隆抗体和角膜上皮细胞特异性标志蛋白K3/K12抗体AE5进行免疫组织化学鉴定。结果 皮肤干细胞在角膜基质的调控下可以横向分化发育成表达角膜上皮细胞特异性标志蛋白K3/K12的细胞 ,呈现AE5抗体阳性 ;并可形成类似角膜上皮外观透明的多层细胞结构。结论 利用皮肤干细胞有可能重建角膜上皮 ,进行自体生物角膜的构建。
Objective To observe the differentiation and development of skin stem cells on corneal stroma and to explore the possibility of reconstructing corneal epithelium by skin stem cells. Methods The primary and secondary cultured human skin and rabbit skin stem cells were cultured on rabbit corneal stroma. The differentiation and development of human skin stem cells were observed in vitro and in vivo. The experimental specimens were stained with HE and identified by immunohistochemistry with human epithelial keratin K19 monoclonal antibody and corneal epithelial cell specific marker protein K3 / K12 antibody AE5. Results Under the regulation of corneal stroma, skin stem cells could differentiate into cells expressing corneal epithelial cell specific marker protein K3 / K12. AE5 antibody was positive, and a multilayered cell structure similar to that of corneal epithelium could be formed. Conclusion The use of skin stem cells may reconstruct corneal epithelium and construct autologous cornea.