Fig. 2

Long LPS exposure significantly increases the therapeutic efficacy of MSC. MSCs were priming in the presence or absence of LPS for 1, 24, and 48 h and coculture with splenocytes that were previously stained with the fluorescent dye CellTrace Violet (CTV) and activated for 72 h with concanavalin A (ConA, 1 μg/ml). a Representative histogram analysis of T cell proliferation by flow cytometry. b Percentage of CD3+ T cells proliferation (%) after coculture with or without MSCs. Data are expressed as mean ± SEM; n = 3, N = 3 biological replicates; *p < 0.05, **p < 0.01. Statistical analysis was performed by one-way ANOVA, Kruskal-Wallis ad hoc post test. c, d EAE in vivo model: MSCs after priming with LPS (500 ng/m for 1 and 24 or 48 h) were injected 7 days after EAE induction. Clinical symptoms and weight loss were evaluated daily. c Clinical score and d cumulative clinical score are shown. Representative data of three independent experiments. Data are expressed as mean ± SEM; n = 12. * symbol represent the comparison between EAE control group and those groups treated with MSCs (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). δ symbol represent the comparison of EAE+MSCs group versus LPS-treated MSCs groups. (δδ < 0.01). Statistical analysis was performed by c two-way ANOVA or d one-way ANOVA, Kruskal-Wallis ad hoc post test