Figure 2.
Comparison of the Effect of EVs Derived from Early-Passage MSCs versus Late-Passage MSCs
(A) Representative scanning electron microscopy (SEM) images of PD15 and P40 MSC-EVs. (B) PD15 and PD40 MSC-EV particle sizes by nanoparticle tracking analysis. (C) PD15 and PD40 MSC-EV particle numbers normalized by cell number. (D) IL-2 and IFN-γ ELISAs with conditioned medium of splenocytes activated by plate-bound anti-CD3 for 24 h with or without EVs from PD15 or PD40 MSCs. (E) IFN-γ and IL-17 ELISAs with conditioned medium of anti-CD3/28-stimulated splenocytes for 72 h with or without PD15 or PD40 MSC-EVs. (F) TNF-α, IL-6, and IFN-γ ELISAs with conditioned medium of LPS (50 ng/ml)-stimulated splenocytes for 24 h with or without EVs from PD15 or PD40 MSCs. (G) IFN-γ ELISA with conditioned medium of splenocytes activated by plate-bound anti-CD3 for 24 h with or without EVs (3 × 109/mL) from early- or late-passage MSCs (#7075 and #6015). IL-6 ELISAs with conditioned medium of splenocytes stimulated with LPS (50 ng/mL) for 24 h with or without EVs (3 × 109/mL) from early- or late-passage MSCs (#6015 and #7075) are shown. (H) RT-PCR assay for spleen IL-6 mRNAs at 5 h after intravenous (i.v.) injection of EVs (1 × 1010 particles/mouse) in LPS-challenged mice (30 μg/mouse; i.v.; n = 5/group). EVs were isolated from PD15 or PD40 MSCs. As controls, PBS and DEX (1.5 mg/kg) were injected. Data are presented as means ± SD (n = 4).; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 by one-way ANOVA with Dunnett’s test.