Effects Of Different External Factors On Epidermal Stem Cells Of Biological Characteristic

PartⅠCultivation and identification of epidermal stem cells in vitroObjective:Isolation epidermal stem cells from the back skin of neonatal SD mice and culture in vitro, check the cells by morphology and specific markers to ensure that the cells is epidermal stem cells.Methods:Successfully cultured the cells, check the CK19, Integrin betal by immunofluorescence staining and check the CD71 and Intergrin Alpha6 by flow cytometryResults:Immunofluorescence Assay results show that both CK19 and Integrin betal were positive, and flow cytometry show the Intergrin Alpha6 was positive and CD71 was negative.Conclusion:We cultured the epidermal stem cells successfully. PartⅡThe effects of different frequencies Low-intensity pulsed electromagnetic fields on proliferation and differentiation of epidermal stem cellsObjective To observe the epidermal stem cells proliferation and differentiation-related genes Myc, beta-catenin changes in the Low-intensity pulsed electromagnetic fields (LIPEMF).Methods Isolated Epidermal stem cells (EpSCs) from the back skin of neonatal SD mice, the EpSCs were divided into experimental and control groups. And the LIPEMF therapy group was divided into three sub-groups to receive 15,50,75 Hz fields. Each group was observed by morphology of cells, immunofluorescence, RT-PCR and immunohistochemical.Results EpSCs by LIPEMF therapy group could increased the Myc gene and beta-catenin gene, there was a statistically significant (P<0.01). The degree of change was different from 15,50,75 Hz fields,50Hz>75Hz>15Hz (P<0.05).Conclusion LIPEMF can induce the differentiation of EpSCs, and different intensity makes a different results. PartⅢNanog andβ-catenin:a new point to EpSCs proliferation and differentiation.Skin tissue homeostasis is maintained by the balanced proliferation and differentiation of certain types of proliferating cells such as epidermal stem cells (EpSCs). The proliferation and differentiation of EpSCs are complex processes and are little known by researchers. This study tried to find the internal relationship between Nanog pathway and wnt/β-catenin pathway in the proliferation and differentiation process of EpSCs. In brief, EpSCs were isolated from rat epidermis and cultured. MTT assay, Western Blotting, PCR (polymerase chain reaction) and Immunocytochemistry were then respectively done during the proliferation and differentiation process of EpSCs. Our results showed that 10-7M neuropeptide Substance P could effectively stimulate proliferation of EpSCs and a possible link exists between Nanog pathway and wnt/β-catenin pathway.