ABSTRACT
Objective To explore the possibility of basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF)combined with striatal conditioned medium promoting the directional differentiation of rat bone marrow mesenchymal stem cells(BMMSCs)into dopaminergie neurons.Methods 1.Separation and culture of BMMSCs:BMMSCs were harvested from healthy adult Wistar rats for serial subcultivation.2.Preparation of Striatal conditioned medium:newborn Wistar rats within 24 hours were selected,and their brain tissues were removed to prepare striatal conditioned medium.3.Induced differentiation of BMMSCs:the 5th passage BMMSCs were collected and pre-induced in low glucose-Dulbecco's modified eagle medium(L-DMEM)containing bFGF and EGF.Twenty-four hours later,pre-induction liquor was replaced with striatal conditioned medium for further induced differentiation.4.Result assessment:the morphological changes of stem cells were observed under inverted phase microscope.The expression of neuron specific enolage(NSE)and tyrosine hydmxylage(TH)were identified by immunocytochemical technique.Results The cell body of rat BMMSCs contracted into round and spindle shape after induction by bFGF and EGF combined with striatal conditioned medium.Partial neuron-like cells with prominence could be found.Immunocytochemieal detection showed that the percentages of NSE and TH positive cells were(72.70±14.81)% and(34.50±15.93)%,respectively.Conclusion BMMSCs can be induced directionally into dopaminergiC neurons by bFGF and EGF combined with striatal conditioned medium in vitro.