Fig. 1
From: Characterizing the secretome of licensed hiPSC-derived MSCs
Immunomodulatory capabilities of hiMSC- and hBMSC-EV preparations. (A) Representative plots for analysis of CD9, CD63 and CD81 expression on hiMSC-EVs by ImageStreamX Flow Cytometry (IFCM). From all events (1st plot from left), only singlets were selected (2nd plot from left) to plot side scatter (SSC) intensities against the fluorescence intensities of CD9+ (PE-labeled), CD63+ (APC-labeled) or CD81+ (FITC-labeled). (B) Average number of CD9+, CD63+ or CD81+ hiMSC-EVs indicated as objects/mL (mean ± SD of three independent EV-preparations). (C) Representative plots for analysis of proportion activated lymphocytes by ImageStreamX Flow Cytometry (IFCM). From all events (plot top left), only singlets (plot top middle) and live cells (plot top right) were selected to distinguish CD4+ (BV785-labeled) from CD8+ (BV650-labeled) cells (plot bottom middle). Subsequently, for both populations fluorescence intensities of CD25+ (PE-Cy5.5-labeled) were plotted against CD54+ (AF700-labeled; plots bottom right and left, respectively). (D) Preparations of hiMSC-EVs (N = 3) and respective unconditioned control media (PM) were used in a multi-donor mixed lymphocyte reaction (mdMLR). After 5 days, cells were harvested, labeled with anti-CD4, anti-CD8, anti-CD25, and anti-CD54, and selected as shown. Preparations of hBMSC-EVs with (active control; N = 3) or without (non-active control; N = 3) immunomodulatory capability were used as controls (graphs show the change compared to untreated controls; *P < 0.05; **P < 0.01; ***P < 0.001)