M-CSF regulatory macrophages promote angiogenesis and GM-CSF macrophages promote inflammation in vitro. Human monocytes were cultured in M-CSF or GM-CSF for 7 days and then were left unstimulated or were stimulated with LPS, LPS + Ado, or LPS + PGE₂ for 24 hours at which point supernatants were collected for further studies. (A) Cell tube formation in HUVECs was observed after 24-hour exposure to supernatants harvested from stimulated M-CSF and GM-CSF macrophages from 1 representative donor (n = 3 donors total). Representative images were captured by brightfield microscopy with “find edges” contrast applied in ImageJ to be able to see the tubes. (B) Total tube length was measured in pixels manually using ImageJ software on various images of HUVECs exposed to supernatants of macrophages from multiple donors (n = 3). **P ≤ .01; ****P ≤ .0001 between M-CSF and corresponding GM-CSF samples; #P ≤ .05; ##P ≤ .01 for M-CSF samples relative to unstimulated (NS) supernatants. (C) The number of nodes, defined as 3 or more tubes originating from 1 point, was counted manually using ImageJ software on various images of HUVECs exposed to supernatants of macrophages from multiple donors (n = 3). **P ≤ .01; ****P ≤ .0001 between M-CSF and corresponding GM-CSF samples; #P ≤ .05; ##P ≤ .01 for M-CSF samples relative to NS supernatants. (D) Levels of inflammatory cytokines, TNF, IL12p40, and GM-CSF, in the supernatants of stimulated macrophages were measured by ELISA (n = 5-9). *P ≤ .05 relative to LPS stimulation alone; #P ≤ .05; ##P ≤ .01 for GM-CSF relative to corresponding M-CSF samples. Error bars represent SEM.