Bone Morphogenetic Protein-2 regulates in vitro osteogenic differentiation of mouse adipose derived stem cells

OBJECTIVE: The aim of this study is to investigate the feasibility and efficiency of Bone Morphogenetic Protein-2 (BMP-2) in regulating in vitro osteogenic differentiation of mouse adipose derived stem cells (ADSCs). MATERIALS AND METHODS: Mouse ADSCs were isolated from adipose tissues of C57/BL6 mice (age of 4-6 w) and cultured. Surface antigens of passage 3 (P3) ADSCs, including CD31, CD34, CD90, CD105 and CD133, were analyzed using flow cytometry. Overexpression of BMP-2 was achieved through gene transfection of ADSCs. In vitro osteogenic differentiation of transfected and non-transfected ADSCs cultured in specific induction media was evaluated by Alizarin Red staining. In addition, expression of osteoblast-specific gene, Runx2, was analyzed by quantitative RT-PCR (qRT-PCR). RESULTS: Abundant ADSCs could be isolated from adipose tissue. P3 ADSCs expressed stem cell-specific molecular markers, CD90 and CD105 but did not express CD31, CD34 or CD133. BMP-2 could efficiently transfect mouse ADSCs. Alizarin Red staining revealed that more calcified nodules were formed in BMP-2 transfected ADSCs. qRT-PCR further confirmed higher level of Runx2 expression in BMP-2 transfected ADSCs (p < 0.05). CONCLUSIONS: BMP-2 can promote in vitro osteogenic differentiation of mouse adipose stem cells.