Objective: To investigate brain-derived neurotrophic factor (BDNF)-induced differentiation of rat bone marrow stromal cells (MSCs) into neuron-like cells in vitro and observe its neuroprotective effect of BNDF on the differentiated cells, which might provide the better seed cells for treatment of nervous system diseases.
Methods: The fifth-passage MSCs were induced by BDNF and 2-mercapto ethanol beta-ME respectively, 1, 3 and 6 h after which the induced neuron-like cells were counted and compared. At 3 h, the neuron-like cells were identified by the immunocytochemical staining, reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting.
Results: The two induced cells both displayed neuronal morphologies with long and multipolar cell projections, but BDNF-induced cells survived for a longer time than beta-ME-induced ones. The results of immunocytochemical staining showed that the two neuron-like cells expressed nestin, neuron-specific enolase (NSE), neurofilament (NF), microtubule-associated protein (MAP-2), but not glial fibrillary acidic protein (GFAP). RT-PCR detected mRNA-positive NSE, NF and MAP-2 in the induced cells, with also mild positive GFAP mRNA. Western blotting identified also NSE expression in these neuron-like cells.
Conclusion: BDNF alone may induce rat MSCs to differentiate into neuron-like cells in vitro, which have longer lifetime to better serve the purpose of transplantation and gene therapy for nervous system diseases.