A preliminary experimental study on dedifferentiation of epidermal keratinocytes into epidermal stem cells induced by protein kinase C inhibitor GF109203X
Abstract：Objective To investigate the feasibility of dedifferentiation of epidermal keratinocytes into epidermal stem cells induced by protein kinase C inhibitor GF109203X. Methods Human primary epidermal stem cells and epidermal keratinocytes were obtained by enzyme digestion method and typeⅣcollagen coated chosen method. The cells were divided into 3 groups as follow: experimental group: primary epidermal keratinocytes were cultured for 2 days before treated with GF109203X, of which the final concentration was 10 μM; negative controlled group: the same volume of DMSO was given instead of GF109203X as in experimental group; positive controlled group: human primary epidermal stem cells cultured in the same period without any intervention except for changing medium every other day. Growth of the cells cultured in vitro was observed by inverted microscope. Monoclonal antibody of integrinβ1, keratin 19 (CK19), CK14, and CK10 were detected by immunocytochemical staining. Results Epidermal stem cells exhibited rapid adherence to typeⅣ collagen, the morphology of the cells was round, which had refractivity and formed distinct clones after 4 days culture, the expressions of integrinβ1, CK19 and CK14 were positive. Epidermal keratinocytes could not adhere rapidly to typeⅣ collagen, the morphology of the cells was close round, with different size, which had no refractivity and formed few clones after 4 days culture, the expression of CK10 was positive. Immunocytochemical staining showed that the expressions of integrinβ1, CK19 and CK14 were positive in experimental group and negative in negative controlled group, but the former exhibited less positive cells after being treated with GF109203X for 2 days, for the both of the two groups, CK10 expression was negative. Positive controlled group showed the opposite result: the expressions of integrinβ1, CK19 and CK14 were negative and CK10 was positive. Conclusion GF109203X may induce the epidermal keratinocytes to convert into epidermal stem cells, which provide a nontransgenic approach for dedifferentiation of epidermal keratinocytes into epidermal stem cells. On the other hand, the result of the study may offer a new idea for dedifferentiation of epidermal keratinocytes into induced pluripotent stem cells (iPSC).
Keywords：Keratinocytes epidermalDedifferentiationProtein kinase C inhibitorEpidermal stem cells
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 132-135 )