The Research On Mechanism Of HaCaT Inducing MSCs To Differentiate Into Epidermal Stem Cells

Objective: To study the mechanism of HaCaT inducing MSCs to differentiate into epidermal stem cells.Methods: 1 The MSCs were isolated, cultured by density gradient centrifugation and the identification of MSCs was by detecting CD34, CD44 and CD45 with immunohistochemistry. 2 In the experimental group, HaCaT and MSCs were co-cultured by using transwell; while in the control group, MSCs was cultured alone as routine method. After 3 day and 7 day, MSCs were identificated by detecting ck19, pan-ck andβ1 integrin with immunohistochemistry. 3 The experiment was divided into three groups, there were groups of co-cultured cells, MSCs and HaCaT respectively. The contents of EGF in supernatant were measured by ELISA method.Results: 1 The cells isolated by Density gradient centrifugation were spindle-shaped. Immunohistochemistry shows the cell was negative for CD34, 45 and positive for CD44. 2 After 3day and 7day of co-culture, Immunohistochemistry revealed that MSCs were positive for ck19, pan-ck andβ1 integrin. The positive rate (ck19, pan-ck andβ1 integrin) of the MSCs co-cultured for 7day were higher than that of MSCs co-cultured for 3day. 3 The EGF level of co-cultured group was higher than that of MSCs and HaCaT group. Conclusion: 1 HaCaT can induce MSCs to differentiate into epidermal stem cells. 2 EGF is one of factors involved in differentiation of MSCs to epidermal stem cells.