Induction and differentiation of rat bone marrow mesenchymal stem cells into neuron-like cells in vitro / 中国组织工程研究

ABSTRACT

BACKGROUND:It is a key to choose an appropriate method to trans-differentiated bone marrow mesenchymal stem cells (BMSCs)into neuron-like cells for clinical therapy of neural system injury.OBJECTIVE:To observe the feasibility of the differentiation of rat BMSCs supplemented with rat dentate gyrus extract(DGE)into neuron-like cells.METHODS:DGE was applied to induce rat BMSC trans-differentiation,using 20,40,60,80 mg/L protein concentration.Neuron differentiation was measured using Western blot to screen an optimal dosage.After trans-differentiation,different concentration treated cells were collected.Morphology change was observed following differentiation under an inverted microscope.Neuron specific enolase(NSE)and NeuN expression was determined using Western blot.NeuN expression in cells was detected using immunofluorescence technique.RESULTS AND CONCLUSION:After 7 days,60 mg/L DGE was the optimal induction dose detected by Western blot.With increased concentration,NSE and NeuN expression was increased.At 80 mg/L mass concentration,NSE and NeuN expression was reduced.At 60 mg/L DGE,BMSCs following induction became long,with synapses,Immunofluorescence NeuN staining found that neuron-like cells were positive for NeuN following induction.Results indicated that DGE is an effective biological inductor that can induce BMSC differentiation.