PPARβ/δ-Interfering Peptide Enhanced Mesenchymal Stromal Cell Immunoregulatory Properties

Figure 5

Role of the interaction between PPARβ/δ and NF-κB TF65 subunit in hAD-MSC immunoregulatory property on macrophages. (a) Experimental design of human macrophage differentiation and activation. CD14⁺ population was sorted from human PBMC using CD14 MicroBeads (Miltenyi) and cultured with 20 ng/ml of M-CSF during 7 days. Then, the resulting macrophages were activated with 100 ng/ml of lipopolysaccharides (LPS) and cultured alone (Mϕ), naïve hAD-MSC (Mϕ+ untretated hAD-MSC) or PP11-pretreated hAD-MSC (Mϕ+PP11 hAD-MSC) at ratio 1 : 10 for 24 h. Cells were harvested to analyze macrophage phenotype by FACS. (b) Representative dot plots showing the M2-like macrophage population. Macrophages were analyzed for the expression profile of M2 markers including CD163 and CD206 to determine the percentage of M2-like macrophages CD163⁺CD206⁺ when LPS-activated macrophages were cultured alone (Mϕ), in the presence of naïve hAD-MSC (Mϕ+ untretated hAD-MSC) or hAD-MSC pretreated with PP11 peptide (Mϕ+PP11 hAD-MSC). (c) Representative dot plots showing the M1-like macrophage population. Macrophages were analyzed for the expression profile of M1 markers including CD86 and HLADR to determine the percentage of M1-like macrophages CD86⁺HLADR⁺. The statistical analysis was performed using the nonparametric Kruskal-Wallis followed by Dunn’s post hoc test for multiple comparisons. The data were obtained with hAD-MSC from 3 donors () on 2 independent experiments. Statistical significance was noted as ns for and for .

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