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. 2021 Sep 1;12(6):1409–1422. doi: 10.14336/AD.2021.0621

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Copyright: © 2021 Kim et al.

this is an open access article distributed under the terms of the creative commons attribution license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Figure 4. — Immunomodulatory effects of EVs from early-passage iMSCs with manipulated expression of TGF-β1 and TGF-β2. (A) The level of TGF-β1 or TGF-β2 in EVs derived from P5 iMSCs transfected with siRNAs against TGF-β1 or TGF-β2 was determined with ELISA. (B) After treatment with EVs (3X10⁹ particles/ml) from control (CON) or TGF-β1/2 knockdown (KD) iMSCs, IFN-γ level in conditioned medium of LPS-stimulated splenocytes (18 hrs) was determined with ELISA. (C) Levels of IFN-γ and IL-2 in conditioned medium of splenocytes activated by anti-CD3 (6 hr) with or without EVs (3X10⁹ particles/ml) were determined with ELISA. (D) The level of TGF-β1 protein in EVs derived from P5 iMSCs transfected with control vector plasmids (ΔCON) or TGF-β1 plasmids (ΔTGFb1) was determined with ELISA. (E) After treatment (18 hrs) with EVs (0.75 to 3 X10⁹ particles/ml) from control or TGF-β1-overexpressing iMSCs at a series of concentrations, IFN-γ level in conditioned medium of splenocytes activated by anti-CD3 was determined with ELISA. All 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.