Optimized protocol for dissociation of mouse intestinal epithelium to viable single-cells

The functional profiling of intestinal epithelial cells (IECs), which comprise multiple heterogeneous populations, is crucial for understanding host interactions with dietary and microbial factors. Here, we present a protocol for dissociating mouse epithelium based on TrypLE Express and Dispase II. We describe steps for tissue collection, crypt isolation, and enzyme digestion. This protocol enables the recovery of millions of viable IECs from the mouse intestine and is particularly useful for isolating rare cell populations, including enterochromaffin (EC) cells.