Induced Differentiation Of Human Bone Marrow Derived Mesenchymal Stem Cells In Vitro Toward Osteoblasts

Objective: To investigate the feasibility of inducing in vitro mesenchymal stem cells ( HMSCs ) derived from human bone marrow differentiate into osteoblasts and potential applicability of the HMSCs as the seed cells in tissue engineering.Methods: Human bone marrow was collected from the fractured patient without blood diseases to obtain the HMSCs, which, after in vitro culture in DMEM supplemented with 20% fetal bovine serum and incubation under standard condition, were induced to differentiate into osteoblasts in DMEM containing beta-glycerophosphate (2nmol/L), ascorbic acid (50ug/ml) and dexamethasone (10nmol/L). Proliferation and differentiation of the BMSCs were observed continually under inverted phase-contrast microscope. alkaline phosphatase (ALP) in the BMSCs by Gomori, the calcified nodules were stained by von Kossa method, and the changes in the content of ALP were measured.Results :The BMSCs proliferated rapidly in in vitro culture and after a 3- to 4-week induction, strong reaction for ALP and calcified nodules were observed. Increasing ALP secretion by the BMSCs was seen as the time of induction prolonged (P<0.05).Conclusions: Human bone marrow-derived HMSCs can be induced to differentiate into osteoblasts through relatively simple procedures, which provide ideal autogenous source of seed cells for bone tissue engineering. The method adopted in this experiment may be used for bone tissue engineering.