BIO (6-bromoindirubin-3'-oxime) GSK3 inhibitor induces dopaminergic differentiation of human immortalized RenVm cells

Abstract

Parkinson's disease (PD) is one of the most neurodegenerative disorders which can lead to severe neural disability and neurological defects. Cell-based therapy using fully differentiated cells is a new method for the treatment of this abnormal condition. In the present study, we investigated the effects of 6-bromoindirubin-3'-oxime (BIO) on dopaminergic differentiation of human immortalized RenVm cells in order to obtain a set of fully differentiated cells for transplantation in Parkinson's disease. To this end, the immortalized RenVm cells were induced to dopaminergic differentiation using a neuro basal medium supplemented with N2 and different concentrations (75, 150, 300, 600, and 1200 nM) of BIO for 4, 8, and 12 days. The efficiency of dopaminergic differentiation was determined using immunocytochemistry for tyrosine hydroxylase expressions. In addition, the expression of a beta-catenin marker was measured using the western blot technique. The results of immunocytochemistry revealed that the mean percentage of Tuj1- and TH-positive sells in 150- and 300-nM-BIO-treated groups was significantly increased compared to that of other groups (p </= 0.01). In addition, the expression of the beta-catenin marker was higher in these groups as compared with that of other groups. Overall, BIO through its effect on the Wnt-Frizzled signaling pathway can promote dopaminergic differentiation of RenVm cells in a dose-dependent manner.