Figure 2.

Conditioning results in an enhancement of the immunomodulatory capacity of EVMSC-T. (a) Peripheral blood mononuclear cells (PBMCs) were stained with CFSE and stimulated with anti-CD3 and anti-CD28 beads in the presence or absence of EVMSC-T or MSC-T^c-derived EVs (EV_MSC-T^c). After 5 days, cells were stained with anti-CD3, anti-CD4 and anti-CD8 antibodies and proliferation of T-cell subsets was determined by flow cytometry measurement of CFSE dilution. Suppression (percentage) was calculated based on the expansion index. Graphs represent mean ± SD of four independent experiments. Unpaired t-test was used for statistics. (b) Representative Western blot of IDO, COX2 and PD-L1 protein expression levels. CD9 was used as a loading control. (c) PD-L1 protein expression level measured by flow cytometry using MFI geometric mean value of EV_MSC-T and EV_MSC-T^c stained with anti-PD-L1. Values are represented as relative to EVMSC-T geometric mean value. Graphs represent mean ± SD of three independent experiments. Paired t-test was used for statistics. * p < 0.05, *** p < 0.001.