Figure 8.

Following persistent stimulation of MSCs with TNFα + IL-1β, the resulting inflammatory CAFs release factors that promote scattering of luminal-A BC cells. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in Figure 1) or to vehicles, generally for 14–18 days. Cytokine-devoid CM were collected as described in Figure 7 and were administered to mCherry-expressing MCF-7 (A) and T47D (B) human luminal-A BC cells, followed by determination of tumor cell scattering out of spheroids (MCF-7 cells) or the ability to form spheroids (T47D cells). (A1,B1) Representative images of spheroid formation at days 8 or 4 for MCF-7 cells and T47D cells are demonstrated, respectively. Bar, 200 µm. The results of a representative experiment out of n = 3 are presented. (A2,B2) Quantitative analyses of spheroid sizes performed by the ImageJ program on images of the representative experiment (n ≥ 7 images for each treatment). *** p < 0.001.