[Inducing human marrow mesenchymal stem cells into osteoblasts directionally and identification of their osteogenesis characteristics].
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.


CONCLUSIONS
We have successfully induced human MSCs into osteoblasts; the induced cells have typical osteogenesis characteristics.
Yin
Xiao-xue
XX
Department of Orthopedic Surgery, Third Hospital of Peking University, Beijing, P. R. China 100083.
Chen
Zhong-qiang
ZQ
Guo
Zhao-qing
ZQ
D000293:Adolescent; D000469:Alkaline Phosphatase / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D002648:Child; D024042:Collagen Type I / Q000096:biosynthesis; D003907:Dexamethasone / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D010012:Osteogenesis; D023822:Tissue Engineering
D024042:Collagen Type I; D003907:Dexamethasone; D000469:Alkaline Phosphatase