Human Bone Marrow Mesenchymal Stem Cells Differentiated Into Dopaminergic Neuron-like Cells In Vitro.

ObjectiveTo explore the potential of human bone marrow mesenchymal stem cells differentiated into neuron-like Cells and dopaminergic neuron-like cells in vitro.MethodsThe human BMSCs were isolated by density gradient centrifugation, and purified by plastic-adhering culture in vitro. Then the purity of BMSCs was analyzed by flow cytometry by detecting CD29,CD44,CD166,CD14,CD34 and CD45. After pretreatment with basic fibroblast growth factor(bFGF) and epidermal growth factor(EGF)48h,then the third passage of BMSCs were cultured at the medium containing glial cell line—derived neurotrophic factor(GDNF) and Angiotensin II(AngⅡ) . The cell morphology of BMSCs during the differentiation was observed by inverted microscope. The expression of Nestin and NSE and TH of BMSCs were detected by RT-PCR in induced group and control group. The expression of NSE and Nestin and TH during the induced group and the control group was detected by IF cytochemistry.ResultsThe third passages BMSCs were homogeneous fusiform shape under the light microscope, the high purity of BMSCs were collected and indicated to express CD29,CD44 and CD166,but not CD34 and CD45. The results showed that 2 weeks after the differentiation of BMSCs by AngⅡthe majority of the BMSCs were transformed into neuron-like cells. There were several processes (axon-like or dendrite-like) from the cell body. The Nestin and NSE mRNAs expressed by BMSCs after induction were more than those before induction significantly (P < 0.05). The result of IF cytochemistry indicate that the expression of NSE,TH increased significantly after induction (P < 0.05), but the GFAP was nagtive whatever before or after induction.ConclusionsThe high purity of BMSCs were gain through the density gradient centrifugation and plastic-adhering culture in vitro, the expression percentage of CD29,CD166and CD44 were above 95%. but no CD34 and CD45. AngⅡcan induce Human BMSCs to differentiate into neuron-like cells in vitro, the neuron-like cells express Nestin and NSE,TH,but not GFAP, Above of all , BMSCs can become the resource of transplantation in neurogenic disease.