TY  - JOUR
TI  - Engineering Decellularized Extracellular Matrix-Incorporated Apical-Out Airway Organoids
AU  - Gong, Zhuowei 
AU  - Ikpechukwu, Chika Sussan
AU  - Bhattaram, Dhruv 
AU  - Ryan, Amy L.
AU  - Weiss, Daniel J.
AU  - Ren, Xi 
VL  - 16
IS  - 17
PY  - 2026
DA  - 2026/09/05
SP  - e5799
C1  - Bio-protocol 2026;16:e5799
DO  - 10.21769/BioProtoc.5799
UR  - https://doi.org/10.21769/BioProtoc.5799
AB  - The airway epithelium interfaces with the external environment through its apical surface and with the extracellular matrix (ECM) through its basolateral surface. To model this organization in vitro, we developed a decellularized ECM-incorporated apical-out airway organoid (dECM-AoAO) platform in which human bronchial epithelial cells (HBECs) self-assemble around human lung-derived decellularized ECM microparticles (dECM-MPs). This configuration preserves apical-out polarity while enabling direct epithelial–ECM interactions. Here, we describe a protocol for the vacuum filtration and quantification of dECM-MPs, the generation of dECM-AoAOs, and ultimately, whole-mount immunofluorescence staining for organoid characterization.
KW  - Decellularized extracellular matrix microparticles (dECM-MPs)
KW  - Apical-out airway organoids (AoAOs)
KW  - Human bronchial epithelial cells (HBECs)
KW  - Airway epithelial differentiation
KW  - Epithelial-ECM interactions
KW  - Whole-mount immunofluorescence
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False