Figure 7.
Comparison of miRNA Profiles of EVs from Early-Passage MSCs, 3D MSCs, and Late-Passage MSCs
(A) Venn diagram of miRNAs expressed in 2D_PD15 EVs, 3D_PD15 EVs, and 2D_PD40 EVs. (B) Top 10 ranked miRNAs in 2D_PD15 EVs. (C) Volcano plot of the differentially expressed miRNAs in 2D_PD15 EVs versus 3D EVs or PD40 MSC-EVs. The y axis indicates the −log 10 of the p values and the x axis is the fold change (FC) (measured as the log2-transformed ratio of the expression between both experimental groups). (D) Confirmation of relative let-7b and miR-21-5p expression in EVs from 2D-cultured PD15 and PD40 MSCs and 3D-cultured PD15 MSCs. (E) IFN-γ, IL-17, and TGF-β1 ELISAs with conditioned medium of anti-CD3/28-stimulated splenocytes for 72 h with or without EVs from PD15 MSCs transfected with target miRNA inhibitor (n = 4). (F) IL-6 ELISA with conditioned medium of LPS-stimulated splenocytes for 24 h with EVs derived from PD15 MSCs transfected with target miRNA inhibitor (n = 4). All data are presented as means ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 by one-way ANOVA with Dunnett’s or Tukey’s test.