Abstract:

Abstract:Objective To explore the method of obtaining and expanding epithelial stem cells (ESCs) from plucked hair follicle in vitro and clarify that fibroblasts are necessary for ESCs growth.Methods A amount of ESCs from plucked hair were obtained,the epithelial outgrowth of human plucked hair follicle was studied and compared between different culture conditions including no coating well,rat tail-collagen coating well,collagen Ⅳ coating well and 3T3 fibroblasts feeder.The characteristics of these cells were detected by RT-PCR and Immunofluorescent staining for CK15.Results After compared between different culture conditions including no coating well,rat tail-collagen coating well,cllagen Ⅳ coating well and 3T3 fibroblasts feeder,the results indicated that the plucked hair fragment B3 (bulge area of hair follicle) placing on 3T3 fibroblasts feeder was the optimum condition.RT-PCR and immunofluorescent method  confirmed that these cells expressed CK15 which was the special marker of ESCs.Conclusion ESCs can be obtained from bulge area of hair follicle which grows on the  3T3 fibroblasts feeder and this method is the optimum condition for ESCs growth after compared between different conditions.

Key words: epithelial stem cells;cell culture;hair follicle