Figure 3.

TGF-β MSCs Establish an Immunosuppressive Microenvironment Unrelated to Nitric Oxide (NO) Production

Untreated or TGF-β MSCs were co-cultured with T lymphocytes at a ratio of 1:10 (MSCs/lymphocytes) for 96 h in the presence of anti-CD3/CD28 polyclonal-stimulating beads. (A–D) Supernatants were collected and centrifuged at 1,000 × g for 5 min to remove cell debris and then analyzed by (A) a Griess assay to measure nitrates, (B) an ELISA to measure PGE2, and (C and D) a magnetic Luminex assay to measure (C) sTNFR1 and sTNFR2 and (D) TNF-α, IFN-γ, granzyme B, and IL-6. Supernatants were used neat. Error bars show mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, by one-way ANOVA, Tukey’s post hoc test (n = 3).