| Part I Induced differentiation of adult HMSCs in vitro toward osteoblastsObjective :To investigate the feasibility of inducing in vitro mesenchymal stem cells derived from adult human bone marrow differentiate into osteoblasts and potential applicability of the MSCs as the seed cells in tissue engineering. Methods: Mesenchymal stem cells were extracted from the bone marrow in blood and cultured immediately in DMEM medium containing 100 m L/L fetal bovine serum. hMSCs were analyzed by the flow cytometry to detect the surface antigens. The subcultured cells was induced to differentiate into osteoblasts with an osteoinductive medium containing dexamethasone (1x10-8 mol/L), beta-sodium glycerophosphate (10 mmol/L) and ascorbic acid (50 mg/L). Cellular morphologies were observed under phase-contrast microscope and the growth curve was drawn accordingly.. The collagen type I and osteocalcin was detected by immunohistochemistry, alkaline phosphatase in the MSCs stained by NAP, the calcified nodules were stained by von Kossa method. Results: The MSCs proliferated rapidly in in vitro culture. Flow cytometry analysis showed that in vitro expanded hMSCs expressed mesenchymal cell marker, including CD105,CD44,and didn't express CD14,CD34,CD 45. The staining of the ALP,COL-I,osteocalcin and calcified nodules were positive after osteoinduction. Conclusions:Human bone marrow-derived MSCs can be induced to differentiate into osteoblasts through relatively simple procedures,.Therefore the MSCs is perfect seeding cells for bone tissue engineering.
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