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Updated: Jul 26, 2025
Establishing Human Lung Organoids and Proximal Differentiation to Generate Mature Airway Organoids
Published on: March 23, 2022
Protocol for generating airway organoids from 2D air liquid interface-differentiated nasal epithelia for use in a
Lisa W Rodenburg1, Isabelle S van der Windt1, Henriette H M Dreyer1
1Department of Pediatric Pulmonology, Wilhelmina Children's Hospital, University Medical Center Utrecht, Utrecht University, Member of ERN-LUNG, Utrecht EA 3584, the Netherlands; Regenerative Medicine Center Utrecht, University Medical Center Utrecht, Utrecht University, Utrecht CB 3584, the Netherlands.
Abstract:
We present a protocol to generate organoids from air-liquid-interface (ALI)-differentiated nasal epithelia. We detail their application as cystic fibrosis (CF) disease model in the cystic fibrosis transmembrane conductance regulator (CFTR)-dependent forskolin-induced swelling (FIS) assay. We describe steps for isolation, expansion and cryostorage of nasal brushing-derived basal progenitor cells, and their differentiation in ALI cultures. Furthermore, we detail the conversion of differentiated epithelial fragments into organoids of healthy controls and CF subjects for validating CFTR function and modulator responses. For complete details on the use and execution of this protocol, please refer to Amatngalim et al.1.
