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. 2016 Apr 13;6:24120. doi: 10.1038/srep24120

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Copyright © 2016, Macmillan Publishers Limited

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(a) Methodological approach to induce MSC priming with EVs (as described in Materials and Methods section). (b) Immunophenotypic analysis of MSCs exposed to resting or primed EVs. IFN-γ/TNF-α-treated MSCs were used as positive control. The histograms show the isotopic controls (dotted curve) and specific markers of control (filled curve) or treated (black curve) MSCs derived from three independent experiments. (c) Resting and primed EVs were incubated with resting MSCs to induce the immunosuppressive phenotypic switch. After 48 hours, sorted-T, -B and –NK cells were added to MSC cultures to assess their immunomodulatory properties. IFN-γ/TNF-α-treated MSCs were used as positive control. CFSE fluorescence was analyzed after 6 days for T and NK, while for B cells the fluorescence was analyzed after 4 days of co-culture. The results are expressed as percentage of relative increase in MSC immunosuppression, normalized to the effect of untreated MSC on IECs (100%). Error bars represented mean ± SEM of five independent experiments. ^*P < 0.05, ^***P < 0.001.