Recapitulating apicobasal tissue polarity in extracellular matrix incorporated airway organoids

Gong, Z.; Bhattaram, D.; Porritt, L.; Golestan, K.; farimani, A. B.; Ryan, A. L.; Weiss, D.; Ren, X.

doi:10.1101/2025.09.03.672699

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bioRxiv · 10.1101/2025.09.03.672699

Recapitulating apicobasal tissue polarity in extracellular matrix incorporated airway organoids

Gong, Z.·Bhattaram, D.·Porritt, L.·Golestan, K.·farimani, A. B.·Ryan, A. L.·Weiss, D.·Ren, X.

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Abstract

The airway epithelium is a dynamic barrier that interfaces with the external environment and internal matrix along its apicobasal axis. To recapitulate this tissue arrangement in an organoid format, we present the decellularized ExtraCellular Matrix-incorporated Apical-out Airway Organoid (dECM-AoAO) that integrates basolateral matrix cues through incorporation of human lung dECM microparticles, while maintaining direct apical exposure to the exterior. Compared to the ECM-free AoAO, dECM incorporation effectively diversifies lineage distribution that better recapitulates native epithelial composition. Harnessing dECM-AoAO locomotion powered by its outward-facing ciliary beating, we developed an experimental and computational pipeline for swarm analysis of organoid group motility as a functional readout of ciliary function. Lastly, dECM-AoAO withstood cryopreservation and revival with sustained viability, lineage composition, and ciliary function, enabling future scalability and broad distribution. Together, this work establishes dECM-AoAO as a more physiologically relevant model system for investigating epithelial-ECM crosstalk during airway homeostasis, pathogenesis, and injury responses.

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Gong, Z., Bhattaram, D., Porritt, L., Golestan, K., farimani, A. B., Ryan, A. L., Weiss, D., Ren, X.. 2025-09-04. Recapitulating apicobasal tissue polarity in extracellular matrix incorporated airway organoids. https://doi.org/10.1101/2025.09.03.672699

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