目的探索人表皮干细胞原代培养方法。方法将皮肤标本进行分离,用DispaseⅡ酶消化表皮皮片后,重悬细胞,接种于铺有Ⅳ型胶原的培养瓶进行培养。表皮干细胞的鉴定采用免疫细胞化学技术检测其表面标记物β1整合素、CK19的表达。对表皮干细胞与角质形成细胞的克隆形成情况进行比较。结果倒置相差显微镜下观察细胞形态及生长状况良好,表皮干细胞β1整合素及CK19角蛋白染色阳性。表皮干细胞克隆形成率高于角质形成细胞(P<0.05)。结论采用本方法进行人表皮干细胞的培养较为理想。 Objective To explore the method of primary culture of human epidermal stem cells. Methods The skin specimens were separated and the epidermal skin pieces were digested with Dispase Ⅱ enzyme. The cells were resuspended and seeded in culture flasks with type Ⅳ collagen. Epidermal stem cells were identified by immunocytochemistry to detect the surface markers β1 integrin, CK19 expression. Clonal formation of epidermal stem cells and keratinocytes was compared. Results The morphology and growth of the cells were observed under inverted phase contrast microscope. The β1 integrin and CK19 keratin staining of epidermal stem cells were positive. The formation rate of epidermal stem cells was higher than that of keratinocytes (P <0.05). Conclusion The method of human epidermal stem cell culture is ideal.