Neural Differentiation of Human Umbilical Cord Blood-derived Mesenchymal Stem Cells

Background: Cell therapy is a potential therapeutic approach for neurodegenerative disorders. Human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) are an appropriate source of stem cells for use in various cell-based therapies. Objectives: In this study, we examined a real-time PCR approach for neural differentiation of hUCB-MSCs in vitro. Materials and Methods: MSCs were cultured in DMEM medium supplemented with 10% FBS in a humidified incubator equilibration at 5% CO2 and 37°C. For the neural differentiation of MSCs, the DMEM was removed and replaced with pre-induction media (retinoic acid [RA], basic fibroblast growth factor [bFGF], and epidermal growth factor [EGF]) and basal medium for two days. They were then cultured in nerve growth factor (NGF), 3-isobutyl-1-methylxanthine (IBMX), ascorbic acid (AA), and basal medium for six days. We also monitored the expression of markers for neural differentiation with real-time PCR. Results: The results of real-time PCR showed that the expression of the GFAP, MBP, and MAP-2 genes after two-step induction was significantly increased compared to the common induction protocol. In addition, our study showed that RA should play the main role in the neural differentiation and fate of MSCs compared to other neural inducers. Conclusions: Taken together, the combination of chemical and growth factors in the two-step induction protocol may improve the efficiency of MSC differentiation into neural progenitor cells, and may be a new method for the easy and fast application of MSCs in regenerative medicine in the future.