Extracellular Vesicles Derived from Three-Dimensional-Cultured Human Umbilical Cord Blood Mesenchymal Stem Cells Prevent Inflammation and Dedifferentiation in Pancreatic Islets

Figure 5

M2 polarization ability of hUCB-MSC-derived EVs in a coculture system of islets and pancreatic macrophages. In a transwell system, 50 islets and macrophages were cultured under three conditions: islets with macrophages without hUCB-MSC-derived EVs (islet w/MΦ), islets with macrophages with 2D hUCB-MSC-derived EVs (islet w/MΦ+2D EVs), and islets with macrophages with 3D hUCB-MSC-derived EVs (islet w/MΦ+3D EVs). (a) Concentrations of IL-1β, IL-18, NLRP3 inflammasome, caspase-1, TNF-α, IL-6, and HMGB1in cocultured hIAPP^+/- islet cells treated with EV produced by 2D- or 3D-cultured hUCB-MSCs were measured using qRT-PCR. The error bars represent the standard deviation of measurements in three separate sample runs (). (b) Glucose-stimulated insulin secretion (GSIS) of hIAPP^+/- mouse islets in each group. GSIS was evaluated using ELISA at 48 h after statin incubation of islets in low (60 mg/dL) and high (300 mg/dL) glucose. The error bars represent the standard deviation of measurements in three separate sample runs (). (c) Flow cytometry analysis of cell surface molecules CD206 and CD80 on macrophages cultured in the presence of EVs. The error bars represent the standard deviation of measurements in three separate sample runs (). (d) Medium concentrations of TGF-β and IL-10 were measured by ELISA. The error bars represent the standard deviation of measurements in four separate sample runs (). Columns, mean; bars, SD. , , , and by one-way ANOVA and Tukey’s posttest.

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