Enhanced primary breast fibroblast invasion in the presence of human macrophages conditioned media. Primary human macrophages were treated +/- estrogen (10-10 M) in 2% charcoal-stripped serum-containing media and 24 hr-conditioned media were collected. Primary human breast fibroblasts were seeded in serum-free media into Matrigel invasions chambers in triplicate. Chemoattractants were 2% charcoal-stripped serum (control), control media + estrogen (10-10 M), macrophage-conditioned media, or conditioned media from macrophages treated with estrogen. (A) 24 hr post-seeding, invaded cells were stained with toluidine blue (top panels), imaged and quantified using NIH Image J software (measurement highlighted in red, bottom panels). (B) Data represent average area measurement of invaded cells +/- standard error (SE) of five independent experiments. *P < 0.01 above control or E2 treatment. ϕ, macrophage conditioned media; E2, estrogen treatment.