Advanced human organoid-on-chip with physiological cellular complexity reveals bidirectional secretion patterns
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. However, epithelial release of immune regulators is understudied. Here, we developed and characterized a human intestinal organoid-on-chip (OoC) model for studying secretion both with proliferation dominated and physiologically relevant cell diversity. 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.