鼠表皮干细胞的分离培养及鉴定

Objective: To explore a method for isolating, culturing and identifying epidermal stem cells of rat. Methods: The epidermal stem cells were isolated and cultured after neutronprotease selectively digested the dermo-epidermal junctions. With observation of the cell growth by inverted microscope and electron microscope. Immunocytochemistry was used to observed the expression of β1-integrin, keratin 15(CK15), keratin 19 (CK19), keratin 5 (CK5) and keratin 10 (CK10). The characteristics of the growth curve was explored, and the cell cycle was determined with flow cytometry (FCM) analysis and the colony forming efficiency (CFE) was studied. Keratinocytes were served as controls. Results: With inverted microscope, the cells were epithelial-like. With electron microscope, many mitotic figures can be observed. Immunohistochemical staining revealed that these cells were positive for β1-integrin, CK15, CK19, CK5 and CK10. The cell cycle profile analysis showed that 48.9% cells were in the S stage. The CFE of the epidermal stem cell was 34.7%. Conclusion: Skin Epidermal stem cells of rats were successfully isolated and cultured in vitro.