Figure 2
Impact of IFN-γ on MSCs’ phenotypic characteristics. Microscopic evaluation of non-primed WJ-MSCs and IFN-γ-primed WJ-MSCs (A). Black arrows indicate the presence of intracellular vesicles in IFN-γ-primed MSCs. Microscopic images were obtained with original magnification 20×, and are presented with scale bars of 50 μm. Total cell number, viability, total ATP, and ADP/ATP ratio measured in non-primed and IFN-γ-primed WJ-MSCs (B). No statistically significant differences were observed regarding the total cell number (p = 0.969), viability (p = 0.232), total ATP (p = 0.832), and ADP/ATP ratio (p = 0.999) between the non-primed and IFN-γ-primed MSCs. The only statistically significant differences were observed regarding the total ATP (p < 0.01) and ADP/ATP ratio (p < 0.01) between the positive control group and the rest of the study groups. Surface marker expression of HLA-DR, -DQ, -DP CD10, CD340, and CD49a of non-primed and IFN-γ-primed WJ-MSCs (C). Indirect immunofluorescence against p38 MAP kinase counterstained with DAPI in non-primed and IFN-γ-primed WJ-MSCs (D). White arrows indicated the presence of p38 MAP kinase P38 MAP kinase stain (green) was co-localized with DAPI stain (blue) in cell nuclei of IFN-γ-primed WJ-MSCs. Microscopic images were obtained with original magnification of 20× and 40×, and are presented with scale bars of 50 and 10 μm, respectively. MLR was conducted either in direct or indirect contact (E). Statistically significant reductions in CB-T cell numbers were observed when co-cultured with WJ-MSCs (both IFN-γ-primed and non-primed) in MLR experiments (p < 0.001). Gene expression analysis, regarding the REX1, OCT4, NANOG, SOX9, KLF4, and GAPDH in non-primed and IFN-γ-primed WJ-MSCS presented in gel electrophoresis (F). Fold change (2−ΔΔCt) of REX1, OCT4, NANOG, SOX9, KLF4and GAPDH expression levels were assessed in non-primed and IFN-γ-primed WJ-MSCs (G). The gene GAPDH was utilized as a housekeeping control for the normalization of gene expression levels. A statistically significant difference in REX1 expression (p < 0.001) was found between non-primed and IFN-γ-primed WJ-MSCs.
