The Effects Of Bone Marrow-derived Endothelial Progenitor Cells On The Proliferation And Differentiation Of Spinal Cord-derived Neural Stem Cells

Objective To investigate the effects of bone marrow-derived endothelial progenitor cells on the proliferation and differentiation of spinal cord-derived neural stem cells.Methods Bone marrow mononuclear cells were isolated by density gradient centrifugation methods and EPCs were cultured by EBM-2 Basal Medium, identified by fluorescent immunocytochemistry. spinal cord-derived NSCs were isolated, cultured and identified by the mature methods. 1×105 /ml tertiary NSCs were plated on the base of culture wells, the upper transwell compartment was seeded with 1×105 /ml primary EPCs, EPCs and NSCs(1∶1) were co-cultured in vitro, set the untreated tertiary NSCs as a control group. At 7 days after co-culture, the number and diameter of neurospheres were calculated and measured with the double blind method, after that, NSCs were maintained for 7 days in DMEM/F12 + 5 % serum medium, and immunochemically stained with neuronal specific marker β-tubulin-Ⅲ, ratio of positiv e cells to total cells was calculated.Results The average number of neurospheres in co-culture group was(22.27±3.85) and the average diameter was( 61.70 ± 7.21) μm, the percentage of neurons immunostaining was(46.10+3.70)%, differences are statistical significance compared with the control group(P<0.01).Conclusion Bone marrow-derived EPCs promoted the proliferation of spinal cord-derived NSCs and induce the differentiation of spinal cord-derived neural stemcells into neurons.