02163nas a2200313   4500000000100000008004100001260001500042653003800057653002100095653002100116653004700137653003200184653001900216100001700235700002000252700001800272700001800290700002500308700001300333700001600346700002000362700001100382245010000393856007200493300001100565490000800576520125100584022001401835       2026                            d  c2026-08-0110aApical-out airway organoid (AoAO)10aCiliary motility10acryopreservation10aDecellularized extracellular matrix (dECM)10aMucociliary differentiation10aSwarm analysis1 aZhuowei Gong1 aDhruv Bhattaram1 aLaura Porritt1 aKian Golestan1 aAmir Barati Farimani1 aBin Deng1 aAmy L. Ryan1 aDaniel J. Weiss1 aXi Ren00aRecapitulating apicobasal tissue polarity in extracellular matrix-incorporated airway organoids  uhttps://www.sciencedirect.com/science/article/pii/S0142961226001080  a1240840 v3313 aThe airway epithelium is a dynamic barrier that interfaces with the external environment and internal matrix along its apicobasal axis. To recapitulate this spatial organization and associated cell-ECM interactions in an organoid format, we present the decellularized extracellular matrix-incorporated Apical-out Airway Organoid (dECM-AoAO), which integrates basolateral matrix cues through the incorporation of human lung dECM microparticles (dECM-MPs), while maintaining direct apical exposure to the exterior. Compared to ECM-free AoAOs, dECM incorporation diversifies lineage distribution, more closely recapitulating native epithelial composition and responsiveness to pathogenic stimulation. Harnessing dECM-AoAO locomotion driven by outward-facing ciliary beating, we developed an experimental and computational pipeline for batch analysis of organoid motility as a functional readout of ciliary activity. Furthermore, dECM-AoAOs are compatible with cryopreservation, retaining viability, lineage composition, and ciliary function upon revival. Together, this work establishes the dECM-AoAO as a physiologically relevant model system for investigating epithelial-ECM crosstalk during airway homeostasis, pathogenesis, and injury responses.  a0142-9612