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Updated: Aug 25, 2026
Isolation of Human Mesenchymal Stem Cells and their Cultivation on the Porous Bone Matrix
Published on: February 9, 2015
[Inducing human marrow mesenchymal stem cells into osteoblasts directionally and identification of their osteogenesis
Xiao-xue Yin1, Zhong-qiang Chen, Zhao-qing Guo
1Department of Orthopedic Surgery, Third Hospital of Peking University, Beijing, P. R. China 100083.
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.
