关键词: bFGF, EGF, BMSCs, 分化, 神经样细胞

Abstract: Objective To investigate the neural differentiation of rat bone marrow mesenchymal stem cells(BMSCs) induced by bFGF and EGF.Methods BMSCs were harvested from Sprague Dawley rats. Flow cytometry was used to assay cell surface marker CD90 and CD45 of BMSCs at passage 3.BMSCs at passage 3 were divided into 4 groups: control group(1% fetal bovine serum + DMEM/F-12), EGF group(20ng/mLEGF), bFGF group(20 ng/mLbFGF) and EGF+bFGF group (20ng/mLEGF+ 20ng/mLbFGF), Then the cellular morphology was observed under inverted phase contrast microscope. Western blot was used to detect neuron specific enolase(NSE) and glial fibrillary acidic protein(GFAP),and RT-PCR used to detect these mRNA.Results BMSCs present a single long spindle or flat-shaped morphology,grow with close whirlpool-like arrangement, the CD90 express up to 98.72%, while CD45 only 1.05%. BMSCs become shrinkage, enhance refraction,display simple bipolar to complex multi-polar, extend distinct processes around cell body,exhibit typical neuron-like morphology after induction.The expression level of NSE and GFAP in group EGF、bFGF and EGF+bFGF was significantly higher than that in control group, significantly higher group bFGF and EGF+bFGF than in group EGF, significantly higher in group EGF than in control group, and significantly higher in group EGF+bFGF than in group bFGF(p<0.0 5).The changes of mRNA was similar to the proteins changes.Conclusion bFGF and EGF are able to promote rat BMSCs to differentiate into neuron-like cells, combined application has the most significant effect.The ability of bFGF to induce BMSCs differentiation into neural cells is stronger than EGF.The present study provide a theoretical basis for the BMSCs transplantation in the treatment of peripheral nerve disorders.

Key words: bFGF, EGF, BMSCs, differentiation, neuron-like cells