Objective: To study the method of inducing human marrow mesenchymal stem cells (MSCs) into osteoblasts directionally and to identify osteogenesis characteristics.
Methods: MSCs were isolated from adult marrow using density gradient separation method and were cultured in conditioned medium containing Dex 10(-8) mol/L, beta-GP 10 mmol/L, and AA 50 micrograms/ml. The MSCs attachment formed soon and passage 3 cells were chosen to check osteogenesis characteristics, including alkaline phosphatase assay with modified calcium-cobalt staining method, type I collagen assay with immunohistochemistry, osteopontin and osteonectin assay with in situ hybridization and calcium nodes assay with Von Kossa staining.
Results: Passage 3 MSCs had typical appearance of osteoblasts and could be passaged continuously till passage 10. The rate of ALP expression was 85%. The expressions of collagen type I, osteopontin and osteonectin were positive and calcium nodes were seen by Von Kossa staining.
Conclusion: We have successfully induced human MSCs into osteoblasts; the induced cells have typical osteogenesis characteristics.
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