Figure 5

Macrophages cultured on CFC or IFC tissue show comparable adherence and appearance. (a) In a newly developed macrophage-tissue assay, macrophages were cultured directly on the aortic tissue surface. Monocytes were separated from human PBMC with MACS CD14 MicroBeads. Monocytes were differentiated to macrophages in vitro for 7 days with M-CSF and seeded directly on the surface (intimal side) of the human CFC or IFC treated aorta. A silicone ring held the tissue punch on the bottom of the culture well to ensure direct contact of macrophages and tissue. After a 2-day co-culture, the tissue punch was either analyzed with scanning electron microscopy (SEM), or the macrophages were harvested and their surface marker expression pattern was analyzed by flow cytometry. (b) Representative SEM pictures of the macrophage-tissue co-culture are depicted. The intima surface of CFC (upper row) and IFC (lower row) aortic tissue are shown with macrophages adhered to the tissue surface (black arrows). In higher magnification (right column) the attached macrophages are visible (black arrows). Scale bars represent 200 µm (left column) and 20 µm (right column).