Journal: Regenerative Therapy

Article Title: Prednisolone attenuates the immunomodulatory capacity and reshapes the functional phenotype of canine adipose-derived mesenchymal stromal cells

doi: 10.1016/j.reth.2026.101184

Figure Lengend Snippet: Effects of prednisolone on macrophage polarization and regulatory T (Treg) cell induction mediated by cADSCs. (a) Representative flow cytometry plots showing cluster designation (CD) 80 and CD206 expression in CD14 + DH82 cells. Cells cultured alone under control conditions or stimulated with interleukin (IL)-4/IL-13 or lipopolysaccharide (LPS)/interferon (IFN)-γ are shown, together with DH82 cells co-cultured with cADSCs pretreated with prednisolone at the indicated concentrations (0–100 μM) under LPS/IFN-γ stimulation. (b) Quantitative analysis of macrophage subsets, defined as CD14 + CD80 + CD206 - , CD14 + CD80 − CD206 + , CD14 + CD80 + CD206 + , and CD14 + CD80 − CD206 - cells. IL-4/IL-13 stimulation was included as an M2-polarizing control, whereas LPS/IFN-γ stimulation induced M1-like polarization. (c) Representative flow cytometry plots showing CD25 and Foxp3 expression in CD4 + T cells after co-culture of PBMCs with cADSCs pretreated with prednisolone. Red boxes indicate CD25 + Foxp3 + Treg cells. (d) Quantification of CD25 + Foxp3 + Treg cells within the CD4 + T-cell population. Data are presented as mean ± SD from three independent experiments using cADSCs derived from three independent donors. * p < 0.05 vs. control or PBMCs alone; † p < 0.05 vs. LPS/IFN-γ.

Article Snippet: To induce macrophage polarization, either a combination of 50 ng/mL lipopolysaccharide (LPS; Sigma-Aldrich) and 20 ng/mL recombinant canine interferon (IFN)-γ (R&D Systems), or a combination of 40 ng/mL recombinant human interleukin (IL)-4 (Thermo Fisher Scientific) and 40 ng/mL recombinant human IL-13 (Thermo Fisher Scientific), was added to the cultures, followed by incubation at 37 °C in 5% CO 2 for 72 h. At the end of the co-culture period, cells were collected, washed with FACS buffer, and incubated on ice with canine Fc receptor binding inhibitor.

Techniques: Flow Cytometry, Expressing, Cell Culture, Control, Analysis, Co-culture Assay, Derivative Assay