Dual adherent/suspension co-culture microenvironment for the study of cancer-stromal interactions using single-cell derived sphere formation

Considerable evidence suggests that cancer-stromal interactions are critical in tumor pathogenesis, but conventional dish-based co-culture methods lack in capability of precisely controlling microenvironments and tracking single cell behavior. To better model tumorigenesis, we present a dual adherent/suspension co-culture device, which can provide both a suspension environment for cancer cells and an adherent environment for stromal cells in close proximity by selectively patterning polyHEMA in indented micro-wells. For investigating cellular heterogeneity, single tumor cells were monitored in a microwell surrounded by stromal cells. As the proof of concept, we have successfully demonstrated the elevated stemness and proliferation rate induced by cancer-stromal interactions.