Fig. 4.

Emergence of spatial structure in tumor/macrophage cocultures. (A) Cocultures with no gradients show no spatial structure as the distribution of macrophages (ΜΦ, labeled in green with CD68) and cancer cells is not significantly different (boxplots). (B) Spatial structure emerges when cocultures are performed in the culture system that mimics the tumor microenvironment. (C and D) Effect of cell migration. (C) Comparison of macrophage migration in normal versus ischemic regions. Rose plots show that macrophages move evenly in all directions [direction vectors are not different from a distribution of random vectors (P > 0.5, modified Rayleigh’s test)]. (D) Mean square displacement (MSD) analysis shows that macrophage movements are subdiffusive. Blue line indicates diffusive movement (slope = 1). (E) Percentage of nonviable macrophages increases over time and with the distance from opening. (Right) Representative visual fields. Macrophages are in green, and nonviable macrophages are in red. ***P < 0.001; NS, not significant. Data were obtained from representative cases. Experiments were repeated at least two times.