Fig. 6.

DMSC effect on Th17 phenotype definition is mediated by soluble factors. DMSC-culture supernatant from one EGF-1-free passage was used as conditioned medium for C57BL/6 spleen cells. Spleen cells were stimulated by anti-CD3 and anti-CD28 antibodies in fresh Click’s medium, in Click’s medium diluted one-half with conditioned medium (50 %), or in whole conditioned medium (100 %). Soluble IL-17 levels were evaluated by ELISA after 3 days under nonpolarizing condition (a), in the presence of IL-6 and TGF-β (b), or after a second round of anti-CD3/anti-CD28 stimulation in the absence of DMSC supernatant (c). Each sample was assayed in quintuplicate and significance was analyzed by t-test; standard error of the means are shown. Results are shown from one representative out of three independent experiments. Conditioned medium was supplemented with 10 % fresh fetal bovine serum before use. *p < 0.05, **p < 0.01, ***p < 0.001. IL Interleukin, Th T helper