Fig. 7

Morphology, distribution and compactness of CRC spheroids cultured with colonic fibroblasts. (A) CRC cells stained with Biotracker 400 Blue and CCD-18Co cells stained with Biotracker 555 Orange (first column), or these cells with the dyes swapped (second column), were mixed at 1:1 proportions. Pictures were taken after 72 h of co-culture. The scale bar represents 75 μm. (B) The compaction of the cancer cells within the spheroids was measured after subtracting the area covered by the fibroblasts, since the fibroblasts formed dense foci that would increase the total spheroid compaction. (C) Percentage of the area covered by cancer cells in spheroids co-cultured with and without fibroblasts. Colours indicate the dye used to label the cancer cells (Biotracker 400 Blue or Biotracker 555 orange). Light blue or orange colours represent the CRC spheroids without CCD-18Co, and dark blue and orange indicate the co-cultures with fibroblasts. *** p-value < 0.01 (two-way ANOVA with dye and fibroblasts as explanatory variables, applied independently for each cell line).