Fig. 4

FL-MSCs demonstrate higher Foxp3⁺ T reg induction capacity in comparison to BM-MSCs. CD3/CD28-activated CD3⁺CD25⁻ effector T cells were co-cultured with BM- or FL-MSCs in 6 different ratios. After 4 days, T cells were collected and the expression of CD25 and Foxp3 was determined in CD4⁺ (a, c) and CD8⁺ (b, d) cells by flow cytometry measurements. Statistical summary dot-plot graphs showing the percentage of CD25⁺Foxp3⁺ iT regs in CD4⁺ (a) and CD8⁺ (b) cells. The first black bar represents the percentage of CD25⁺Foxp3⁺ cells in the activated T cells control group (n = 18 for CD4 and n = 12 for CD8 group). Further bars depict the percentage of CD25⁺Foxp3⁺ cells in activated T cells co-culture with either BM-MSCs (red) or FL-MSCs (blue) (n = 13 for CD4 and n = 9 for CD8 conditions). Representative FACS dot plots of CD25⁺Foxp3⁺ iT reg populations in activated T cells control group or in activated T cells co-cultures with either BM-MSCs or FL-MSCs (c, d). Cells were first gated on CD4⁺ (represented in c) or CD8⁺ (represented in d) cells and then the percentage of CD25⁺Foxp3⁺ double positive cells were determined. The flow cytometry representatives of T reg induction assay were selected from 1:6 MSC to T cell ratio. Data are represented as mean value ± SEM collected from 3 different experiments. One-way ANOVA analysis was performed to generate P values. ns, non-significant; Beads, anti-CD3 and anti-CD28 activation beads; TCs, T cells; T regs, regulatory T cells. *P < .05, **P < .01, ***P < .001, ****P < .0001