Study Of CNTF Combines RA Induced Myoblasts Neural Like Differentiation And Correlation With REST

ObjectiveTo investigate the possibility of neural like differentiation of myoblasts induced by ciliary neurotrophic factor(CNTF)and retinoic acid(RA),and whether or not this induction process have the correlation of neuron restrictive silencing factor(REST).MethodsCNTF induced C2C12 dedifferentiation and then RA induced C2C12 neural like differentiated.Experimental grouping and treatment: induction group(dedifferentiation induced by CNTF medium for 3 days,induction of neuronal differentiation on RA medium for 2 days,nerve cell culture medium for 3 days)and control group(culture medium without cntf for 3 days,without RA culture medium for 2 days,nerve cell culture medium for 3 days).Identification and correlation analysis of dedifferentiated and differentiated nerve cells were performed with the cell morphology,immunofluorescence,flow cytometry analysis,Western blot.Compared the expression of REST before and after neuronal differentiation.ResultsAfter co-induced,cells was "cluster-like" growth,some clumps can be suspended growth,the typical "neurosphere"-like performance.Immunofluorescence staining showed that neurospheres could be labeled by neural markers such as NSE,SOX1,GFAP.The neurospheres grew well after replated,and expressed nestin.Western blot showed that CNTF inducedmyoblast differentiation related protein myogenin and P21 expression was significantly.After neural differentiation NSE,SOX1,GFAP expression was significantly increased,and muscle cell specific markers Desmin and REST expression decreased significantly.Flow cytometry showed that the induction group and the control group were significantly different,and the induction group specific positive cell rate was about 84%.ConclusionsCNTF combines RA can induce neural like differentiation of myoblasts,the process may change associated with the expression of REST.