Intestinal organoid co-culture protocol to study cell competition in vitro

Intestinal organoid cultures are a powerful tool to study epithelial cells in vitro, as they are able to proliferate and differentiate into all cell lineages observed in vivo. Co-culturing organoids with distinct genetic backgrounds provides an excellent approach to study contact dependent and independent interactions between healthy and mutant epithelial intestinal cells. Here, we provide 2D and 3D approaches to mouse organoid co-cultures using fluorescently labeled organoids and demonstrate the analysis of these co-cultures using flow cytometry and microscopy-based approaches. For complete details on the use and execution of this profile, please refer to van Neerven et al., 2021.