Differentiation of human bone marrow mesenchymal stem cells into osteoblasts in vitro / 中国组织工程研究

ABSTRACT

BACKGROUND: Cells grew slowly in a low number after conventional induction. Growth factor can accelerate induction speed, but growth factor is expensive. OBJECTIVE: To verify the efficiency of differentiation of human bone marrow mesenchymal stem cells (BMSCs) into osteoblasts in vitro in a inductor without growth factor. DESIGN, TIME AND SETTING: The control cytology observation experiment was conducted from July 2007 to February 2008 at the Center Laboratory of Hospital Affiliated to Medical College of Qingdao University. MATERIALS Bone marrow was collected from diabetic patients undergoing autologous stem cell transplantation. Informed consent was obtained from patients. METHODS: Using density gradient method, BMSCs were separate and primary cultured. At the third passage, BMSCs were incubated in 0.1 ?mol/L dexamethasone, 10 mmol/L ?-glycerine sodium, and 50 ?mol/L vitamin C. BMSCs in the control group were left intact. MAIN OUTCOME MEASURES: Morphology of BMSCs was observed under a phase contrast microscope. Morphological changes were measured by alkaline phosphatase staining and calcified nodules staining after induction. RESULTS: Under the phase contrast microscope, the third passage of BMSCs was uniform, spindle. At 7 days, cells were confluence. Some cells were overlapped over time. Mesenchymal tissues gradually piled up, with mineralization, multiple nodes. At 14 days, brown mineralized nodes were found under an inverted microscope. At 21 days, mineralization in fragmentis were formed. Von Kossa method demonstrated black calcified nodules and positive alkaline phosphatase. CONCLUSION: BMSCs can be induced into osteoblasts easily, with high and rapid proliferation.