Fig. 3

Optimization of 3D coculture model. OVCAR8-DsRed2 and NCI/ADR-RES-EGFP cells were plated at a 1:1 ratio to final seeding cell densities of 1000–5000. Fluorescence signal from cells was recorded up to 12 days and plotted as fold change from day 1 for 1000 (a), 2000 (b), and 5000 (c) cell seeding densities. Representative longitudinal imaging for spheroids with 2000 seeded cells are shown (d). Next, fold change in OVCAR8-DsRed2 fluorescence was divided by fold change in NCI/ADR-RES-EGFP fluorescence to calculate the cell growth ratio (e). Total killing controls (5% bleach) were included, and viability is plotted as a function of fluorescence (f) and CellTiter-Glo® Cell Viability Assay (g). Representative images of total killing controls are shown (h). Scale bar = 1000 μm. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001