Objective：To establish protein expressing profile of human bone marrow mesenchymal stem cells（hMSCs） which were induced into osteoblasts through proteomic and bioinformatic techniques and to further elucidate the precise molecular mechanism of hMSCs osteoblast differentiation. Methods：The total protein extracts were obtained with cell lysis buffer from hMSCs which were induced into osteoblasts. Two-dimensional gel electrophoresis（2-DE） and 2-DE gel analysis software were used to recognize and test protein spots. Clearly expressed protein spots were selected and were identified and analyzed by matrix-assisted laser desorption ionization-time of flight mass spectromety（MALDI-TOF-MS） and bioinformatics． Results：Protein expressing maps of hMSCs induced into osteoblasts were set up. Seventy-seven proteins were identified by MALDI-TOF-MS and bioinformatics，which were classified by Gene Ontology according to molecular function and biological process. Conclusion：Protein expressing maps of hMSCs induced into osteoblasts are built，which will provide basal information to illuminate the molecular mechanism of hMSCs differentiation into osteoblasts and to build protein expression database of osteoblast differentiation from hMSCs.