Figure 6

Up-take of EVs by murine tubular epithelial cells. (A) Representative micrograph of incorporation of EVs collected from HLSCs stained with Dil (red) by mTEC after five hours of incubation. Original magnification: ×630. Nuclei were stained with Hoechst. Three experiments were performed with similar results. (B) Quantification of proliferation rate by BrdU incorporation assay. mTEC (4,000 cells/well) were treated with vehicle, EV, CM and iEV derived from 8,000 HLSCs. Experiments were conducted in triplicate. Data are expressed as mean ± SD; ANOVA with Dunnet’s multicomparison test was performed: *P <0.05 EV versus vehicle. (C) Evaluation of apoptosis of mTEC (25,000 cells/well) treated with vehicle, CM, EV and iEV derived from 50,000 HLSCs by Muse Annexin V & Dead Cell Assay. Data are expressed as mean ± SD; ANOVA with Newmann-Keuls multicomparison test was performed: *P <0.05 EV and CM versus vehicle; #P <0.05 CM versus EV. ANOVA, analysis of variance; BrdU, 5-bromo-2′-deoxy-uridine; CM, conditioned medium; EV, extracellular vesicles; HLSCs, human liver stem cells; iEV, inactivated extracellular vesicles; mTEC, murine tubular epithelial cells; SD, standard deviation.