The role of BMP-7 in chondrogenic and osteogenic differentiation of human bone marrow multipotent mesenchymal stromal cells in vitro.
This study addresses the role of bone morphogenetic protein-7 (BMP-7) in chondrogenic and osteogenic differentiation of human bone marrow multipotent mesenchymal stromal cells (BM MSCs) in vitro. BM MSCs were expanded and differentiated in the presence or absence of BMP-7 in monolayer and three-dimensional cultures. After 3 days of stimulation, BMP-7 significantly inhibited MSC growth in expansion cultures. When supplemented in commonly used induction media for 7-21 days, BMP-7 facilitated both chondrogenic and osteogenic differentiation of MSCs. This was evident by specific gene and protein expression analyses using real-time PCR, Western blot, histological, and immunohistochemical staining. BMP-7 supplementation appeared to enhance upregulation of lineage-specific markers, such as type II and type IX collagens (COL2A1, COL9A1) in chondrogenic and secreted phosphoprotein 1 (SPP1), osteocalcin (BGLAP), and osterix (SP7) in osteogenic differentiation. BMP-7 in the presence of TGF-beta3 induced superior chondrocytic proteoglycan accumulation, type II collagen, and SOX9 protein expression in alginate and pellet cultures compared to either factor alone. BMP-7 increased alkaline phosphatase activity and dose-dependently accelerated calcium mineralization of osteogenic differentiated MSCs. The potential of BMP-7 to promote adipogenesis of MSCs was restricted under osteogenic conditions, despite upregulation of adipocyte gene expression. These data suggest that BMP-7 is not a singular lineage determinant, rather it promotes both chondrogenic and osteogenic differentiation of MSCs by co-ordinating with initial lineage-specific signals to accelerate cell fate determination. BMP-7 may be a useful enhancer of in vitro differentiation of BM MSCs for cell-based tissue repair.
Shen
Bojiang
B
Orthopaedic Research Institute, Department of Orthopaedic Surgery, St. George Hospital, University of New South Wales, Sydney, Australia.
Wei
Aiqun
A
Whittaker
Shane
S
Williams
Lisa A
LA
Tao
Helen
H
Ma
David D F
DD
Diwan
Ashish D
AD
D050156:Adipogenesis / Q000187:drug effects / Q000502:physiology*; D000464:Alginates / Q000494:pharmacology; D000469:Alkaline Phosphatase / Q000378:metabolism; D000943:Antigens, Differentiation / Q000502:physiology; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology; D055419:Bone Morphogenetic Protein 7 / Q000378:metabolism* / Q000494:pharmacology; D002113:Calcification, Physiologic / Q000502:physiology; D018929:Cell Culture Techniques; D019070:Cell Lineage; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000187:drug effects / Q000502:physiology*; D020723:Glucuronic Acid / Q000494:pharmacology; D006603:Hexuronic Acids / Q000494:pharmacology; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D039902:Multipotent Stem Cells / Q000166:cytology / Q000502:physiology; D010012:Osteogenesis / Q000187:drug effects / Q000502:physiology*; D011509:Proteoglycans / Q000096:biosynthesis; D055758:SOX9 Transcription Factor / Q000502:physiology; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology
D000464:Alginates; D000943:Antigens, Differentiation; D055419:Bone Morphogenetic Protein 7; D006603:Hexuronic Acids; D011509:Proteoglycans; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D053782:Transforming Growth Factor beta3; D020723:Glucuronic Acid; D000469:Alkaline Phosphatase