Consistent Osteoblastic Differentiation of Human Mesenchymal Stem Cells with Bone Morphogenetic Protein 4 and Low Serum

• Published in: Tissue engineering. Part C, Methods Methods, 2011-03, Vol.17 (3), p.249-259
• Main Authors: Cordonnier, Thomas, Langonné, Alain, Sohier, Jérôme, Layrolle, Pierre, Rosset, Philippe, Sensébé, Luc, Deschaseaux, Frédéric
• Format: Article
• Language: English
• Subjects: Adult; Aged; Aged, 80 and over; Alkaline Phosphatase - metabolism; Animals; Bioengineering; Biomaterials; Bone marrow; Bone Morphogenetic Protein 4 - pharmacology; Bone morphogenetic proteins; Cell differentiation; Cell Differentiation - drug effects; Cell Differentiation - genetics; Cell Proliferation - drug effects; Ceramics - pharmacology; Flow Cytometry; Gene Expression Profiling; Gene Expression Regulation - drug effects; Genetics; Humans; Life Sciences; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - drug effects; Mesenchymal Stromal Cells - enzymology; Mice; Middle Aged; Morphology; Osteoblasts; Osteoblasts - cytology; Osteoblasts - drug effects; Osteoblasts - metabolism; Osteogenesis - drug effects; Osteogenesis - genetics; Physiological aspects; Proteins; Serum - metabolism; Stem cells; Tissue engineering; Tissue Scaffolds - chemistry
• ISSN: 1937-3384; 1937-3392
• DOI: 10.1089/ten.tec.2010.0387

Providing fully mature and functional osteoblasts is challenging for bone tissue engineering and regenerative medicine. Such cells could be obtained from multipotent bone marrow mesenchymal stem cells (MSCs) after induction by different osteogenic factors. However, there are some discrepancies in results, notably due to the use of sera and to the type of osteogenic factor. In this study, we compared the osteogenic differentiation of bone marrow MSCs induced by dexamethasone (Dex) or bone morphogenetic proteins (BMPs) by assessing phenotypes in vitro and functional osteoblasts in vivo . Reducing the content of fetal calf serum from 10% to 2% significantly increased the mineral deposition and expression of osteoblastic markers during osteogenesis. In comparison to Dex condition, the addition of BMP4 greatly improved the differentiation of MSCs into fully mature osteoblasts as seen by high expression of Osterix. These results were confirmed in different supportive matrixes, plastic flasks, or biphasic calcium phosphate biomaterials. In contrast to Dex-derived osteoblasts, BMP4-derived osteoblasts from MSCs were significantly able to produce new bone in subcutis of nude mice in accordance with in vitro results. In conclusion, we describe a convenient ex vivo method to produce consistently mature functional osteoblasts from human MSCs with use of BMP4 and low serum.