Advanced human organoid-on-chip with physiological cellular complexity reveals bidirectional secretion patterns

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Intestinal epithelial cells are essential for maintaining gut homeostasis, serving as a physical barrier separating luminal content from the immune compartment. They are crucial for balancing tolerance to commensal and protection from pathogenic bacteria and dietary antigens by the asymmetric secretion of immune regulators. To understand how different epithelial cell types and external stimuli shape this secretion pattern, we established and characterized a human intestinal organoid-on-chip (OoC) model. OoCs were used to evaluate barrier integrity and cell type-dependent expression profiles by RNA-sequencing and microscopy. Secretome analysis revealed asymmetric bidirectional secretion and a cell type-dependent response to LPS, flagellin, gliadin, and cytokines. In summary, we have developed an OoC model using human intestinal cells, offering a cutting-edge platform to study barrier function under physiological and pathophysiological conditions. This versatile tool holds significant promise for advancing our understanding of epithelial cells in innate immunity and driving the development of personalized therapeutic strategies.

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