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

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

  • STAR Protoc. 2023 Jun 12;4(3):102337. doi: 10.1016/j.xpro.2023.102337.
Lisa W Rodenburg 1,  Isabelle S van der Windt 1,  Henriette H M Dreyer 1,  Shannon M A Smits 1,  Loes A den Hertog-Oosterhoff 1,  Ellen M Aarts 1,  Jeffrey M Beekman 2,  Gimano D Amatngalim 3
Affiliations

Affiliations

  • 1 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.
  • 2 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. Electronic address: [email protected].
  • 3 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. Electronic address: [email protected].
PMID: 37314920 DOI: 10.1016/j.xpro.2023.102337
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.

Keywords

Cell culture; Organoids.

Figures
Products