Crosstalk with Inflammatory Macrophages Shapes the Regulatory Properties of Multipotent Adult Progenitor Cells

Figure 4

Soluble factors derived by macrophages-primed rMAPC modulate macrophages phenotype. (a) Schematic illustration of application of rMAPC-derived conditioned media to macrophages (MΦ). (b) TNF-α ( independent experiments with triplicates per experiment) and (c) IL-6 ( experiments with triplicates per experiment) secretion levels of macrophages treated with LPS in the presence of rMAPC-derived conditioned media. The results are presented as percentage of to the positive control (medium +LPS). Asterisks () indicate statistical significant difference with positive control. (d–h) NR8383 mRNA expression of M1 and M2 polarization markers seeded in DCM ( experiments). iNOS (d), CD86 (e), and TNF-α (f) are compared to their respective positive control (NR8383 +LPS) while Arg1 (g) and CCL18 (h) to NR8383 +M2 cytokines (IL-4/IL-10/IL-13). The results are presented as fold differences to nonstimulated NR8383 cells (dotted line). (i-j) NR8383 mRNA expression of Arg1 seeded in DCM supplemented with polarization stimulus, +LPS (i) and +M2 cytokines (j). The results are presented as fold differences to nonstimulated NR8383 cells (dotted line). Asterisks () indicate statistical significant difference with positive control in each case (NR8383 +LPS or NR8383 +M2 cytokines). Relative expressions were normalized against the expression of hydroxymethylbilane synthase (HMBS) and b-actin. Mean values ± SEM. Data were analyzed with one-way ANOVA followed by Dunnett’s multiple comparison test or with Kruskal-Wallis test followed by Dunn’s multiple comparison test for nonparametric data. Two group comparisons were made with unpaired Student’s t-test (, , and ).

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)

(j)