Fig 3. In vitro generation of canine monocyte-derived macrophages.

Monocytes were obtained by positive selection using CD14 monoclonal antibody coated columns. CD14⁺ monocytes were then seed in 12 or 24 wells plates at 1x10⁶ cell/mL in DMEM enriched with 50 ng/mL of canine GM-CSF. Macrophages were allowed to mature for 7 days upon which were used for the experiments. Flow cytometry after monocyte isolation: (A) Control and (B) CD14⁺ cells. (C) Morphology of adherent macrophages after 7 days of incubation with DMEM and GM-CSF by light microscopy. Scale bar = 20 μm. Similar data were obtained in three independent experiments using macrophages from different animals.