Differentiation of human bone marrow mesenchymal stem cells into dopaminergic neuron-like cells induced by angiotensinⅡ combined with growth factor

Abstract:

Objective：To explore the potential of human bone marrow mesenchymal stem cells（hBMSCs） to differentiate into neurons and dopaminergic neuron-like cells in vitro. Methods：The hBMSCs were isolated and purified from normal human bone marrow mononuclear cells in vitro. After the identification and pretreatment with basic fibroblast growth factor（bFGF） and epidermal growth factor（EGF），the hBMSCs were cultured at the medium containing glial cell line-derived neurotrophic factor（GDNF） and Angiotensin Ⅱ（AngⅡ）. The cell morphology of hBMSCs during the differentiation was observed by inverted microscope. The mRNA expressions and proteins of Nestin，neuron-specific enolase（NSE），human glial fibrillary acidic protein（GFAP），tyrosinehydroxylase（TH） of hBMSCs were detected by RT-PCR and immunofluorescence method before and after the induction. Results：Ｔhe hBMSCs with high purity of were collected. The majority of the hBMSCs were transformed into neuron-like cells displaying as bipolar，multipolar and cone in sharp at two weeks after the differentiation of hBMSCs by AngⅡ. The mRNAs expressions of Nestin，NSE and TH by hBMSCs after the induction were significantly more than those before the induction（P<0.05）. Immunofluorescence analysis showed that the number of cells expressing proteins of NSE，Nestin and TH was increased significantly after the induction（P<0.05）. GFAP gene was not expressed by cells before and after the induction. Conclusion：AngⅡcombined with other growth factors can induce hBMSCs to differentiate into dopaminergic neuron-like cells.