Abstract: OBJECTIVE To investigate the effect of vitexin on osteogenic differentiation of rat bone marrow mesenchymal stem cells(MSCs). METHODS Methylthiazolyl tetrazolium(MTT) assay was applied to determine the viability of MSCs. Alkaline phosphatase(ALP) activity was detected using p-nitrophenyl phosphate assay. Nodule formation of calcium was observed with alizarin red stain. The genes expression of runt-related transcription factor 2 (Runx2) and osterix (Osx) was detected by real time PCR. RESULTS Vitexin at concentration from 0.1 to 50 μmol·L^-1 had no effect on viability of MSCs. Vitexin effectively stimulated the activity of ALP in MSCs and increased the nodule formation of calcium. Vitexin also up-regulated the expression of Runx2 and Osx mRNA. CONCLUSION Vitexin improves the osteogenic differentiation of MSCs, which may be related with the up-regulation of the expression of Runx2 and Osx.