Protocol for generating airway organoids from 2D air liquid interface-differentiated nasal epithelia for use in a functional CFTR assay

Lisa W. Rodenburg; Isabelle S. van der Windt; Henriette H.M. Dreyer; Shannon M.A. Smits; Loes A. den Hertog - Oosterhoff; Ellen M. Aarts; Jeffrey M. Beekman; Gimano D. Amatngalim

STAR Protocols (Sep 2023)

Protocol for generating airway organoids from 2D air liquid interface-differentiated nasal epithelia for use in a functional CFTR assay

Lisa W. Rodenburg,
Isabelle S. van der Windt,
Henriette H.M. Dreyer,
Shannon M.A. Smits,
Loes A. den Hertog - Oosterhoff,
Ellen M. Aarts,
Jeffrey M. Beekman,
Gimano D. Amatngalim

Affiliations

Lisa W. Rodenburg: Department 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
Isabelle S. van der Windt: Department 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
Henriette H.M. Dreyer: Department 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
Shannon M.A. Smits: Department 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
Loes A. den Hertog - Oosterhoff: Department 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
Ellen M. Aarts: Department 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
Jeffrey M. Beekman: Department 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; Centre for Living Technologies, Alliance TU/e, WUR, UU, UMC Utrecht, Utrecht CB 3584, the Netherlands; Corresponding author
Gimano D. Amatngalim: Department 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; Corresponding author

Journal volume & issue: Vol. 4, no. 3
p. 102337

Abstract

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Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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