Isolation and culture of rhesus putative epidermal stem cells / 中国病理生理杂志

ABSTRACT

AIM: Our study was designed to look for an easy and feasible approach to isolate and culture rhesus epidermal stem cells.METHODS: Skin specimens were cut into strips and immersed into 0.25% trypsin overnight.Then transparent cuticular layeres were striped off with ophthalmic microscopic forceps.The epithelial layers were scratched,harvested and transferred to culture in skin epithelium media.The cells in 2nd-4th passages were harvested by 0.25% trypsin and relayed in IV collagen plate at 37 ℃,5% CO_2 for 20 min to harvest rapid attaching cells.Flow cytometric analysis,immunohistology and RT-PCR were conducted to identify rapid attaching cells.RESULTS: The specific protein and mRNA of epidermal stem cells(?6 integrin,K15 and ?1 integrin) were identified in rapid attaching cells.No K1/K10(marker of terminally differentiated cells) and CD71 expression were found.CONCLUSION: Our method of isolation and culture can apply for rhesus epidermal stem cells.