FIGURE 1.

Cytokine preconditioning of MSCs (p6) in a SF‐medium (SF+CYT) causes the upregulation of the immunomodulatory markers and preserves their stemness potential. A, MSCs were committed toward osteogenic (O) or adipogenic (A) lineages after the preconditioning protocol. Calcium deposits (O, in red) are visualized by Alizarin Red, while fat droplets (A, in red) are stained by Oil Red, indicating osteocytic and adipocytic differentiation, respectively. CTRL‐O and CTRL‐A: controls of MSCs grown in the absence of osteogenic (O) and adipogenic (A) inducing differentiation media. Images were acquired by phase contrast microscopy. Magnification ×20. Scale bars = 50 μm. B, Immunoblotting evaluation of the expression of typical stemness (CD90 and CD73) and immunoregulatory markers (COX2 and IDO) by MSCs subjected to SF + cytokine (SF+CYT) or SF preconditioning (CTRL: control; SF: serum‐free; CYT: cytokines, TNFα and IFNγ). C, Histograms relative to the quantification of the immunomodulatory marker bands in (B). For the comparison between groups (SF+CYT at 24 and 48 hours) unpaired, two‐tailed Student's t test was used. All the data are expressed as mean ± SEM (n = 3). MSC, mesenchymal stem cell; SF, serum‐free