Fig. 1
Morphology of MSC treated with CM. a Experimental scheme used for generating conditioned medium (CM) from macrophages and for MSC treatments. CMGM+ and CMM+ were obtained from MΦGM and MΦM, respectively, that were treated with LPS for 90 min, thoroughly washed with PBS, and further incubated in fresh medium for 5 h. CMGM− and CMM− were obtained from MΦGM and MΦM, respectively, that were not treated with LPS but subjected to identical experimental manipulations. MSC were incubated or not with CM for 48 h, thoroughly washed with PBS, and further incubated in growth (GrM) or osteogenic (OM) medium up to 21 days. b Optical microscope images showing the morphology of MSC treated or not (−) with CM after crystal violet staining (upper row). Confocal microscope images (maximum projection) showing actin (red) and nuclei (blue) in MSC treated or not with CM (lower row). c Ratio of cell length to width as measure of cell elongation. *p < 0.05 between the indicated conditions