Complex of neural stem cells and endothelial progenitor cells enhances differentiation of neural stem cells to neuron and formation of synapse after implanted in MCAO rats / 第三军医大学学报

ABSTRACT

Objective To explore the effect of the complex containing neuron stem cells(NSCs)and endothelial progenitor cells(EPCs)after being transplanted to ischemia and reperfusion rat model on the differentiation of neural stem cells to neuron and synapse formation.MethodsTotally 300 SD adult male rats were randomly divided into sham operation group,middle cerebral artery occlusion(MACO)model group,MACO+NSCs group,MACO+EPCs group,and MACO+complex group,and every group were further divided into 6 subgroups according to time points 1,3,7,14,30 and 60 d after operation,NSCs were isolated from the hippocampus of neonatal SD rats born in 24 h and identified with Nestin staining.EPCs were obtained from the artery blood of SD rats and verified with immunocytochemical stainings of CD31 and CD34 after culture.NSCs and EPCs were cultured together to produce the complex with aid of laminin.Then normal saline,the NSCs,EPCs,or complex(2?1010/L)were transplanted into the ischemia penumbra of corresponding model rat brain,sham control or MACO rats.The differentiation of NSCs,the expression of P38,synapsin-I,mRNA of connexin 43 were observed with double label immunofluorescence technique,immunohistochemical method,RT-PCR and Western blot analysis in ischemic penumbra.ResultsCompared with other groups,there were more double Brdu and neural special enolase positive cells in MACO+complex group with the increased expression of P38.RT-PCR and Western blot analysis indicated that MACO+complex group had significantly higher expressions of synapsin-I at mRNA and protein levels in ischemic penumbra,and so did the mRNA expression of connexin 43.ConclusionThe transplantation of the complex of NSCs and EPCs improves the NSCs differentiation to neuron,synapse formation and reconstruction of neural network.