Notch Signaling Is Involved In Neuronal Differentiation Of Rat Tendon-Derived Stem Cells

Objective:To explore the neuronal differentiation potential of tendon-derived stem cells(TDSCs)in rat Achilles' s tendons in vitro,test the expression changes in the related genes of neuronal cells,and study the regulation mechanism and Notch pathway in differentiation process.Methods:The TDSCs were isolated from Achilles tendon of the 4-week-old SD rats,and cultured to the third generation(P3).The control group continued the normal culture without any treatment,while medium of the experimental group were replaced by the neuronal-differentiation medium.Real-time quantitative polymerase chain reaction(Real-time PCR),Western blot(Western Blot)and immunofluorescence techniques were used to detect the expression changes of nerve cell-related genes GFAP,Neun and ?3-tubulin.Also the expression changes of Notch pathway-related genes,NICD,Hes-1 and RBPJk were detected.The change of the ability of TDSCs to differentiate into neuronal cells was detected after regulation of Notch pathway.Results:Compared with the control group,the morphology of TDSCs after induction of neuronal differentiation changed,which trended to the morphology of nerve cells,and the expression of nerve cell-specific genes increased at the gene,protein and cell levels.The expression of related genes of Notch pathway were decreased during the differentiation process.Compared with normal TDSCs,the ability to differentiate into neuronal cells was increased after Notch pathway was inhibited.Conclusion:TDSCs isolated from rat have the ability to differentiate into neuronal-like cells,and the Notch pathway is involved in the process.Notch pathway is inhibited during the differentiation.The ability of neuronal differentiation of TDSCs can be enhanced by inhibiting Notch pathway.The study can provide a theoretical basis for the clinical study of chronic pain caused by neoinnervation in tendinopathy.