Establishment Of The Model Of Marrow Mesenchymal Stem Cells Differentiation In Vitro And Its Application To Mechanism Investigation On Bone Damage Caused By Environmental Chemicals

Objectives to establish and optimize in-vitro culture system of murine mesenchymal stem cells (BMSCs) in order to obtain more BMSCs; to establish the model of BMSCs differentiation in vitro; to investigate the mechanism of adverse effects on BMSCs by sodium fluoride and alcohol and therefore to provide theoretical basis for the prevention and cure of osteofluorosis, osteoporosis and osteonecrosis by alcohol abuse.Methods 1 the marrow was harvested from C57/BL6 male mice. BMSCs were amplificated in vitro by the method of whole marrow culture and identified surface marker CD90 and CD34 by immunohistochemistry. Finally the growth curve of BMSCs was described. 2 BMSCs were cultured in a-MEM supplemented by dexamethasone, β-sodium phosphoglycerol and vitamin C to differentiate toward ostoblasts. Cells were harvested at different time to be identified the early, middle and late function markers of osteoblasts. Furthermore BMSCs were induced in a-MEM supplemented by dexamethasone and insulin to differentiate toward adipocytes. Cells were obtained to conduct oil...