Abstract:

BACKGROUND: Wnt signaling pathway is a key regulator of cellular proliferation and differentiation, but its correlation with neural differentiation of bone marrow mesenchymal stem cells (BMSCs) is not very clear.

OBJECTIVE: To find out the molecules of the Wnt family which are involved in differentiation of rat BMSCs into neuron-like cells.

METHODS: The rat BMSCs were separated and cultured in vitro. The morphology of the BMSCs was observed. Flow cytometry analysis was performed to detect cell phenotype CD44, CD9, CD34 and CD45. Wnt3a and Wnt5a were respectively combined with basic fibroblast growth factor to induce BMSCs differentiation into neuron-like cells, and then were identified by using immunocytochemistry and RT-PCR.

RESULTS AND CONCLUSION: The BMSCs were long-spindle. CD9 and CD44 were highly expressed, while CD34 and CD45 were lowly expressed. Nestin and neuron specific enolase were positive but glial fibrillary acidic protein were not obviously expressed when they were cultured with Wnt3a. In Wnt5a group, Nestin expression was weakly positive, while neuron specific enolase and glial fibrillary acidic protein were negative. RT-PCR result revealed Nestin expressed both before and after induction in the Wnt3a induced group, neuron-specific enolase exhibited apparent amplified bands 5 days after the induction, and more apparent at 10 days. A weak amplification band of glial fibrillary acidic protein could be seen at 10 days after the induction. In Wnt5a and control groups, BMSCs induced by 10 days weakly expressed Nestin, while neuron-specific enolase and glial fibrillary acidic protein were almost not expressed. It is indicated that Wnt3a molecule can promote the differentiation of BMSCs cultured in vitro to neuron-like cells.