Objectives To induce the differentiation of rabbit bone marrow mesenchymal stem cells (MSCs) to myogenic cells in vitro, and to investigate the expression of vascular endothelial growth factor (VEGF) gene in MSCs transfected by AdTrackCMV-VEGF165. Methods MSCs were isolated and purified from rabbit bone marrow by percoll (1073 g/ml) and then cultured in low glucose DMEM with 10% FBS. AdTrackCMV-VEGF165 eukaryotic expression vector was constructed and transfected into the MSCs. After being incubated with 5-azacytidine (5-Aza), the expression of troponin I in MSCs was assayed by immunohistochemistry and the expression of VEGF gene was identified by northern blot and western blot.The concentration of VEGF in the supernatant was measured by ELASA. Results MSCs were isolated and cultured successfully from rabbit bone marrow. The positive cTnI stain of some MSCs after the induction of 5-aza indicated that the cells were differentiated to myogenic cells. Northern blot and western blot showed that the expression of VEGF 165 mRNA was much higher in the hVEGF165 gene transfected cells than that of the control. The concentration of VEGF in the supernatant got to the peak 3-5 days after hVEGF165gene transfection (1011- 1027 pg/ml) and decreased gradually thereafter, but still higher than that of control group or pAdTrackCMV group (349 pg/mL vs 116 pg/mL or 125pg/ml respectively, P＜ 0.01). Conclusions MSCs could be induced to differentiate to myogenic cells by 5-aza in vitro and could express VEGF by VEGF gene transfection.