Fig. 5
EVs produced in the hollow-fiber system from hBM-MSCs produce consistent glycan profiles. Glycan analysis of EVs isolated from each hBM-MSC donor (N = 4 donors; hBM-MSC-48RB/55RB/81RB/85RB) over several days (days 1, 13, and 25) confirms consistent glycoprofiles of EVs. a, b Lectin microarray analysis of EVs (66.7 μg/mL) labeled with DyLight-555-NHS. a Heat map showing the relative intensity of fluorescently labeled EVs, from four individual hBM-MSC donors at three collection time points, bound to 26 glycan-binding lectins on a microarray slide (Z-Biotech). b Quantitative analysis of the lectin microarray fluorescence signal of the mean of the four hBM-MSC donors at the three collection time points is represented. Each dot represents N = 4 donors, each analyzed in triplicates. Statistical significance was analyzed by a two-way analysis of variance (ANOVA) with Tukey’s post hoc analysis. A p value of < 0.05 was considered statistically significant and significant differences are marked with a single (p < 0.05), triple (p < 0.001), or quadruple (p < 0.0001) asterisks. c Qualitative comparison of the capillary electrophoresis-laser-induced fluorescence (CE-LIF) traces from enzymatically released and APTS-labeled glycan profile from EV samples collected at three collection time points during EV production from four hBM-MSC donors. The traces for the glycan profile for the EV samples harvested at day 1 (red), day 13 (black), and day 25 (blue) are displayed for donors hBM-MSC-48RB (i), hBM-MSC-55RB (ii), hBM-MSC-81RB (iii), and hBM-MSC-85RB (iv), respectively