Osteogenic Differentiation of Human MSCs

MSCs were trypsinized and placed in a 12-well plate or a 24-well plate at a density of 4000 cells/cm². The cells were incubated in α-MEM medium for three or four days. When over 80% confluence was reached, the medium was then replaced by an osteogenic induction medium (OIM) containing 1 nM dexamethasone, 20 mM beta-glycerophosphate, and 50 μM ascorbic acid. MSCs were treated for two weeks. The osteogenic differentiation was evaluated by real time qRT-PCR after 11 days and Alizarin Red staining at day 3, day 5, day 7 and day 11. Triplicate tests were conducted in each experiment.