CEP supplemented in 3D fibrin matrix increases macrophage migration, but not neutrophil migration. Thrombin-treated fibrinogen forms a 3D polymerized gel in a Boyden chamber. Thioglycollate-induced WT (A) or β2−/− (B) peritoneal macrophages were labeled with PKH67 green fluorescent dye and plated on the gel. Macrophage migration was stimulated with 30 nM MCP-1. A total of 9 µM EP (left) or CEP (right) was incorporated in the gel during polymerization. Results were evaluated in 4 to 6 samples per group (9 field of view per sample), analyzed by IMARIS 8.0 software, and plotted. (C) Neutrophils were labeled with PKH26 red fluorescent dye and plated on fibrin matrix with incorporated CEP or EP. The migration was detected after 24 hours as described for macrophages. The first 30 μm of the gel from the starting point (where many nonmigrated cells reside) is not shown to reduce a gradient of brightness intensity for the sample. Statistical analyses were performed using Student paired t tests (n = 4 per group). Bar represents 500 μm.
