Figure 7.

Enhanced Mϕ proliferation by hAM-MSCs via CCL2 and TGF-β1 pathways

Mouse bone marrow-derived Mϕ were cultured with or without hAM-MSCs in a non-contact co-culture model for 48 h. (A) Phase-contrast images showing the Mϕ frequency. Mϕ were dissociated for cell number counts, the results of which are presented in the chart. Scale bars, 400 μm. n = 6. (B) Immunocytolabeling showing Ki67 expression in Mϕ. Collected Mϕ were stained for a proliferation marker Ki67 and DAPI, and percentage of Ki67⁺ nuclei was measured and present in the chart. Scale bars, 50 μm. n = 4 in each group. (C) Effects of inhibition of human CCL2 and TGF-β1 on hAM-MSC-mediated Mϕ proliferation. Increased Mϕ numbers by hAM-MSC co-culture were eliminated by addition of neutralizing antibodies for hCCL2 (Mϕ + hMSC + anti-CCL2 group) and TGF-β1 (Mϕ + hMSC + anti-TGF-β1 group). Isotype (IgG) antibody used as control. Representative phase-contrast images and a chart summarizing the data are presented. Scale bars, 400 μm. n = 4. Data are presented as mean ± SEM. ∗p < 0.05 and ∗∗p < 0.01. Student’s t test (A and B) or one-way ANOVA with Tukey’s post hoc test (C).