Fig. 7
Relationship between cell density and differentiation. A MSCs were seeded at four concentrations (5 × 102, 5 × 103, 5 × 104, 5 × 105 MSCs/ml) into culture wells. B Multipotent differentiation of MSCs related to cell density. (a) Adipogenic differentiation of MSCs. The graph shows quantification of PPAR-γ-positive cell numbers as differentiated adipocytes from independent experiments (means ± SD). (Bar = 40 μm) (b) osteogenic differentiation of MSCs. The graph shows quantification of Runx-2-positive cell numbers as differentiated osteoblasts from four independent experiments (means ± SD). (Bar = 50 μm) C schematic of in vitro coculture study. D Relationship between MSCs and OECs in coculture. (a) Quantification of MSCs number in the upper Transwell insert chamber to determine MSC migration through the 8-μm pores. (Bar = 20 μm) (b) OEC adhesion assay. Data show the percentage of OECs under MSC coculture. Bars represent the means ± SD of four independent experiments. *p < 0.05 (c) scratch assay. OECs were cultured for 3 days then observed by immunofluorescence staining for actin filaments (red). The white dotted line shows the limit of the wound area into which the cells migrated, while the white arrow indicates migrating cells. (Bar = 20 μm) the right panel graphically shows the number of migrating cells under each condition. Bars represent the means ± SD of four parallel experiments. *p < 0.05