Optimized human intestinal organoid model reveals interleukin-22-dependency of paneth cell formation

Opposing roles have been proposed for IL-22 in intestinal pathophysiology. We have optimized human small intestinal organoid (hSIO) culturing, constitutively generating all differentiated cell types while maintaining an active stem cell compartment. IL-22 does not promote the expansion of stem cells but rather slows the growth of hSIOs. In hSIOs, IL-22 is required for formation of Paneth cells, the prime producers of intestinal antimicrobial peptides (AMPs). Introduction of inflammatory bowel disease (IBD)-associated loss-of-function mutations in the IL-22 co-receptor gene IL10RB resulted in abolishment of Paneth cells in hSIOs. Moreover, IL-22 induced expression of host defense genes (such as REG1A, REG1B, and DMBT1) in enterocytes, goblet cells, Paneth cells, Tuft cells, and even stem cells. Thus, IL-22 does not directly control the regenerative capacity of crypt stem cells but rather boosts Paneth cell numbers, as well as the expression of AMPs in all cell types. [Display omitted] •Optimized culture protocol for human intestinal organoids•Organoids display extensive budding and harbor all cell types•IL-22-mTOR signaling mediates human Paneth cell differentiation•IL-22 induces expression of host defense genes in all cell types Generating a long-term organoid culture model that displays extensive budding and harbors all cell types of the human small intestine reveals mechanism of human Paneth cell differentiation by IL-22-mTOR signaling.