Figure 8.

Enhanced migration of monocytes and Mϕ by hAM-MSCs through CCL2

(A–D) Migration ability was assessed in freshly isolated mouse bone marrow-derived monocytes (A and C) or Mϕ (B and D) by seeding in the upper well of a Transwell insert and measuring movement of these cells toward a confluent monolayer of hAM-MSCs (hMSC group) or no cell (media group) in the lower side of the membrane. Effects of co-culture with hAM-MSCs with human CCL2 neutralizing antibody (hMSC + anti-CCL2 group) and administration of recombinant CCL2 with or without anti-CCL2 antibody (CCL2 + anti-CCL2 and CCL2 groups) were also assessed. Migrated cells through the insert membrane were fixed and stained with DAPI and counted. Representative DAPI-staining images (A and B) and respective counts of migrated cell numbers are presented in the charts (C and D). Scale bars, 50 μm. n = 4−5 independent experiments. Conditions were repeated in duplicate for each biological replicate, and an average >5 frames per insert were captured. Data are presented as mean ± SEM. ∗p < 0.05 and ∗∗p < 0.01. One-way ANOVA with Tukey’s post hoc test for multiple paired comparisons.