Neural Differentiation of USSC by Inhibition of GSK-3β

Vahid Dastjerdi F *, Zeynali B , Tafreshi A.P , Soleimani M ,


Objective: Somatic signaling pathway. While there are large body of evidence for the involvement of this GSK-3β is a key molecule in several signaling pathways including canonical Wnt pathway in the right balance between proliferation and differentiation states of the stem cells, none is available on the newly introduced stem cell called the unrestricted stem cell (USSC). Materials and Methods: In this study, we have therefore investigated if the activation of Wnt pathway is involved in the differentiation of USSCs into dopaminergic neurons. USSCs were isolated from umbilical cord and cultured in the presence of neural diffrentiation factors and specific GSK-3β inhibitor (BIO) for 10 days. The culture media of the control USSCs contained neural differentiation factors and the DMSO, a solvent for BIO. The expression of general neuronal marker (β-tubulin III) were examined by using flowcytometery. The activation of Wnt pathway was examined at the expression levels of pGSK-3β and β-catenin by immunocytochemistry and western blotting. Results: Our results showed that in cells cultured in the presence of both neural differentiation factor and BIO, the expression of general neural marker were significantly increased compared to that in the control cells. Moreover, BIO increases the experssion of pGSK-3β and β-catenin compared to that of the control, indicating that Wnt pathway is activated in these cells. Conclusion: Altogether, these result show that the inhibition of GSK-3β in USSCs may lead to neural differentiation suggesting a role for canonical Wnt signaling pathway in differentiation of USSC stem cells.