Abstract:

BACKGROUND: In vitro induced bone marrow mesenchymal stem cells (BMSCs) differentiated into neuron-like cells mainly by polypeptide preparations such as nerve growth factor. Pure chemical inducer is not commonly found.
OBJECTIVE: To establish a system for isolation and culture of BMSCs and induce it directional differentiation into neuron-like cells in vitro.
METHODS: BMSCs were isolated, purified and identified by density gradient centrifugation, adherent culture and digestion time control. BMSCs were induced into neuron-like cells by β-mercaptoethanol and dimethyl sulfoxide. The morphology change was observed. The differentiation ratio analysis of differentiated neuron-like cells was detected by Nissl staining and immunochemical of neuron specific enolase and neurofilament-200.
RESULTS AND CONCLUSION: Separated BMSCs were fibroblast-like cells and they had several nucleoluses. BMSCs could differentiate into neuron-like cells, with the presence of long axons and dendrite-like processes after being induced by β-mercaptoethanol and dimethyl sulfoxide. Both Nissl’s body, neurofilament -200 and neuron specific enolase of neurons after induction were positive and the positive rates of neuron specific enolase and neurofilament-200 were (85.6 ±6.7)% and (73.2 ± 5.6)%, respectively. Results demonstrated that density gradient centrifugation, adherent culture and digestion time control can successfully isolate and culture human BMSCs. Human BMSCs can differentiate into neuron-like cells induced by β-mercaptoethanol and dimethyl sulfoxide in vitro.