Fig. 2

Human T-cell proliferation is suppressed following 24 h exposure to MSCs in a dose-dependent manner. a Schematic of 24 h efferocytic-licensing of monocytes in suspension followed by replating and 4d T-cell activation. b Gating strategy for T-cell proliferation analysis. After singlet discrimination, CFSE positive PBMCs were gated, and the T-cell population was gated using FSC-SSC. % proliferation was then calculated using a gate based on the No Stim Control. c Representative T-cell proliferation histograms for MSC and HI-MSC culture conditions at each MSC:PBMC ratio tested. The dotted line denotes the proliferative (+) from the non-proliferative (−) cells. d Quantification of T-cell proliferation as a percent of the positive stim control for each MSC:PBMC ratio. (Mean ± SEM, n = 3 independent experiments, *p < 0.05 calculated after 2-way ANOVA with Dunnett post hoc test to compare + MSC and + HI-MSC at each ratio to normalized stimulated control (dashed line)). e Quantification of pro- and anti-inflammatory cytokines detected after T-cell activation without or with efferocytic-licensing by viable or HI-MSC (box and whisker show 25th, 50th, and 75th percentile while whiskers show min and max values, n = 3 independent experiments, *p < 0.05 calculated after 1-way ANOVA with Tukey post hoc test to compare all conditions)