Abstract: Objective：To study the in vitro biologic characteristics of bone marrow stromal cells with cleidocranial dysplasia (BMSCs-CCD), including osteogenesis, proliferation ability, stemness and senescence. Methods：MTT and cell cycle detection for proliferation ability, Western blot and alizarin red staining for osteogenesis ability, Western blot and clony-formation for stemness ability, Western blot and senescence staining for senescence characteristics. Results：The optical density value was lower in BMSCs-CCD in MTT experiment and lower BMSCs-CCD were under S or G2 stage while more under G1 stage in cell cycle detection. BMSCs-CCD expressed lower levels of Runx2, Opn, Osterix and weaker alizarin red staining. Moreover, BMSCs-CCD also exhibited weaker stemness associate proteins (Oct4, Nanog, and Sox2) and clony-formation rates. However, stronger senescence associated proteins (p16, p21) and more senescenced cells were found in BMSCs-CCD. Conclusions：BMSCs-CCD exhibited weaker proliferation ability, osteogenesis and stemness while stronger senescence ability as compared to BMSCs. These results are helpful for us to understand pathological mechanism of CCD bone disease.