Organoids as a Model for Intestinal Ion Transport Physiology

The advent of intestinal organoid culture in 2009 was a fortuitous development in the search for a valid marker of intestinal stem cells, and provided proof of murine intestinal stem cell regenerative potential. Intestinal organoid culture was preceded by key discoveries of the Wnt/β-catenin signaling pathway and the development of 3D culture matrices. The latter, involving a laminin-rich gel to provide an artificial basement membrane, was instrumental to primary intestinal epithelial culture by preventing anoikis, an immediate apoptotic event when intestinal epithelial cells detach from the basement membrane. One of the first physiological studies using 3D murine “mini-gut” structures showed cystic fibrosis transmembrane conductance regulator (CFTR) expression and anion channel activity in the crypt-like structures projecting from the epithelial-lined central cavity. Detailed investigations of ion transport physiology using human intestinal organoids, both primary and iPSC-derived, found close similarities to existing knowledge of ion transport physiology and included the development of the forskolin-induced swelling assay (FIS). The FIS assay using organoids cultured from rectal biopsies of cystic fibrosis patients provided an avenue for personalized medicine to test small-molecule modulators on different CFTR mutations. More recent research has led to the development of 2D primary intestinal epithelial monolayers, which provide easy access to the apical, lumen-facing membrane and the opportunity for traditional ion transport studies with Ussing chambers. Human 2D primary intestinal monolayers also demonstrate the dominance of CFTR in anion secretion and provide a quantitative evaluation of its chloride and bicarbonate secretory conductances. These aspects of ion transport physiology using 2D and 3D intestinal cultures are discussed along with the relative advantages and disadvantages of each culture method with respect to technical aspects and recapitulation of native intestinal epithelium.