Schwann Cells Differentiation Process Neuregulin 1/ErbB Pathways And Sox10 Relations Research

Objective This experiment using stem cells have the characteristics of the multi-directional differentiation potential,using in vitro induced bone marrow mesenchymal stem cells to differentiate between schwann's cells,and based on the differentiation process of Neuregulin 1 / Erb B signaling pathways to influence and significance of Sox10 expression.Methods With high sugar separation mice after BMSCs in the generation of culture medium,when fusion cells reached 30%,use add 1 mm beta mercaptoethanol DMEM culture cell 24 h,with added 10% fetal bovine serum and then 35 ng/m L all trans retinoic acid(RA)72 h DMEM culture of cells,cell differentiation treatment before end.The experimental group was divided into three groups.Inducer group: using the classic induction method,the cells were cultured in DMEM with 10%FBS,5ng/m L platelet derivatives(pdgf-aa),10ng/m L alkaline fibrogrowth factor(b FGF),14 m M Forskolin and200ng/m L Heregulin.Blockers(TAK-165),with 10% FBS,5 ng/m L(PDGF AA)and platelet derived factor 10 ng/m L alkaline fibroblast growth factor(b FGF),14 mm Forskolin,50 microns TAK-165(Erb B2 blockers)and 200 ng/m L Heregulin DMEM culture cell,thereby blocking Neuregulin 1 / Erb B pathways.HRG off group: the DMEM culture cells of 10%FBS,5ng/m L platelet derivatives(pdgf-aa),10ng/m L basic fibroblast growth factor(b FGF)and 14 m M Forskolin were added.The expression changes of S100,p75,GFAP,Sox10,neuregulin1,Erb B2 and Erb B3 were detected by real-time fluorescence quantitative PCR.S100,p75 and GFAP were measured by immunofluorescence staining to determine the extent of stem cell differentiation into schwann cells.CCk-8 was used to detect the proliferation of cells in each group.Results Immunofluorescence staining showed positive expression of S100,p75,GFAP in the cells of the inducible group and blocker group.PCR results showed that the expression of S100,p75 and GFAP in the blocker group were significantly lower than that in the induction group,and the difference was significant(P<0.05);Real-time fluorescence quantitative PCR results showed that the expression of neuregulin-1 was consistent and stable after the induction of 0d,1d,4d and 7d,and no significant difference was expressed at all time points(P>0.05),The expression of neuregulin-1 in the HRG off group and the blocker group was not significantly affected(P>0.05),indicating that neuregulin-1 was involved in the whole process of inducing differentiation;Real-time fluorescent quantitative PCR results showed Erb B2 have always had a steady high expression,the differentiation of 0 d,1 d,4 d,7 d when the expression of no significant change(P>0.05),while the Erb B3 does not produce at the beginning of induction,and with the extension of induction time expression gradually rise,explain Neuregulin 1 /Erb B3 signaling pathways role in the differentiation process gradually enhanced;In the process of extracorporeal induction,Sox10 also has a stable expression.Real-time fluorescence quantitative PCR and PCR gel were used to detect the expression of Sox10 in the differentiation of 0d,1d,4d,and 7d,without significant difference(P>0.05),indicating that it was continuously stable during the differentiation process;Compared with revulsant group,blockers in the expression of Sox10 decline,and have significant difference(P<0.05),whereas Sox10 no significant decline in HRG off group(P>0.05),illustrate Neuregulin 1 / Erb B signaling pathway to maintain or promote the expression of Sox10;Among the three groups of cells,the most proliferative capacity was the least differentiated HRG off group,followed by the inducer group,with the lowest proliferative capacity of the TAK165 group,and the difference was significant(P<0.05).Conclusion 1.Neurogulin-1 /Erb B2 signal pathway can promote differentiation of mouse BMSCs into Schwann-like cells.2.It has been proved that Neuregulin-1/ErbB signaling pathway plays an important role in the promotion and maintenance of Sox10 expression,and this effect may further affect the differentiation of bone marrow mesenchymal stem cells into Schwann like cells..