Researchers have developed an iPSC-derived small intestine-on-chip that mimics the human small intestine's structure and function. This system uses organ-on-chip technology to create a more physiologically relevant in vitro environment, allowing for apical and basolateral access to the intestinal epithelial barrier. By exposing cells to growth factor gradients along the crypt-villus axis, the system enables self-organization of epithelial, mesenchymal, and neural cells into villus-like folds with physiological barrier integrity. The result is a well-characterized hiPSC-derived intestine-on-chip system that can facilitate personalized studies on physiological processes and therapy development in the human small intestine. The system includes a diverse epithelial composition and myofibroblast and neural subtypes, which self-organize into villus-like folds in the top channel and emerge in the bottom channel. The growth factor gradients efficiently balance dividing and mature cell types, inducing an intestinal epithelial composition resembling the human adult small intestine. The system also shows resemblance to the human small intestine, with cell-type composition and functionalities that align with those found in human tissue. The intestine-on-chip system can be used to model selective cellular responses to external factors, such as cytokines. Dividing epithelial, mesenchymal, and neural subtypes were shown to be more responsive to IFN-β and IFN-γ stimulation than non-dividing cells, indicating differences in IFN sensitivities between cells in the intestine-on-chip. The system provides a sustainable and reproducible method to model the human small intestinal epithelial barrier, with physiologically relevant cellular diversity, including myofibroblast and neural cell types. The composition of EM and DM includes compounds that activate or inhibit well-established pathways in intestinal development and were previously demonstrated to be effective in controlling epithelial lineage induction in adult stem cell-derived intestinal epithelial organoids. The same differentiation factors used in adult stem cell-derived intestinal organoids can efficiently induce mature lineages in hiPSC-derived intestinal epithelial cells, and even in mesenchymal and neural cells.Researchers have developed an iPSC-derived small intestine-on-chip that mimics the human small intestine's structure and function. This system uses organ-on-chip technology to create a more physiologically relevant in vitro environment, allowing for apical and basolateral access to the intestinal epithelial barrier. By exposing cells to growth factor gradients along the crypt-villus axis, the system enables self-organization of epithelial, mesenchymal, and neural cells into villus-like folds with physiological barrier integrity. The result is a well-characterized hiPSC-derived intestine-on-chip system that can facilitate personalized studies on physiological processes and therapy development in the human small intestine. The system includes a diverse epithelial composition and myofibroblast and neural subtypes, which self-organize into villus-like folds in the top channel and emerge in the bottom channel. The growth factor gradients efficiently balance dividing and mature cell types, inducing an intestinal epithelial composition resembling the human adult small intestine. The system also shows resemblance to the human small intestine, with cell-type composition and functionalities that align with those found in human tissue. The intestine-on-chip system can be used to model selective cellular responses to external factors, such as cytokines. Dividing epithelial, mesenchymal, and neural subtypes were shown to be more responsive to IFN-β and IFN-γ stimulation than non-dividing cells, indicating differences in IFN sensitivities between cells in the intestine-on-chip. The system provides a sustainable and reproducible method to model the human small intestinal epithelial barrier, with physiologically relevant cellular diversity, including myofibroblast and neural cell types. The composition of EM and DM includes compounds that activate or inhibit well-established pathways in intestinal development and were previously demonstrated to be effective in controlling epithelial lineage induction in adult stem cell-derived intestinal epithelial organoids. The same differentiation factors used in adult stem cell-derived intestinal organoids can efficiently induce mature lineages in hiPSC-derived intestinal epithelial cells, and even in mesenchymal and neural cells.