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Objective To investigate the biological characteristics of rabbit bone-marrow mesenchymal stem cells after transfection with Ad-BMP-2/GFP. Methods The 10th generation of rabbit BMSCs were isolated and purified by density gradient centrifugation combined with adherent culture method. Cell surface antigen expression of CD44, CD45, and CD29 was detected using flow cytometry. The cells were transferred by adenovirus carrying BMP-2/GFP gene. The change of cell morphology was observed using an inverted fluorescence microscope. Cell proliferation was analyzed using MTT method. The osteoblast differentiation was studied using immunohistochemical staining for collagen type I and Alizarin red staining for calcium nodules. Targeted protein expression before and after the transfection was detected using Western blot detection. Results The 10th generation of rabbit BMSCs with high purity were successfully obtained using density gradient centrifugation and adherent culture method. Flow cytometry show ed CD44 and CD29 were positive while CD45 was negative. Green fluorescence was observed in rabbit BMSCs under the fluorescence microscope after Ad-BMP-2/GFP transfection. The morphology of the cells show ed osteogenic differentiation. The transfection promoted proliferation of BMSCs in a short time (P < 0. 05). And both type I collagen immunohistochemical staining and Alizarin red staining show ed positive. Western Bloting results indicated that target protein BMP-2 expressed stably in the cells. Conclusion Rabbit BMSCs can be successfully transfected with Ad-BMP-2/GFP and the biological characteristics of the cells are significantly changed after transfection. [ABSTRACT FROM AUTHOR]