Protocol for generating airway organoids from 2D air liquid interface-differentiated nasal epithelia for use in a functional CFTR assay
Publication date
2023-09-15
Authors
Rodenburg, Lisa W.
van der Windt, Isabelle
Dreyer, Henriette H M
Smits, Shannon M A
den Hertog-Oosterhoff, Loes A.
Aarts, Ellen M
Beekman, Jeffrey M.
Amatngalim, Gimano D.
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Advisors
Supervisors
Document Type
Article
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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, General Biochemistry,Genetics and Molecular Biology, General Immunology and Microbiology, General Neuroscience, Journal Article
Citation
Rodenburg, L W, van der Windt, I S, Dreyer, H H M, Smits, S M A, den Hertog-Oosterhoff, L A, Aarts, E M, Beekman, J M & Amatngalim, G D 2023, 'Protocol for generating airway organoids from 2D air liquid interface-differentiated nasal epithelia for use in a functional CFTR assay', STAR protocols, vol. 4, no. 3, 102337, pp. 1-28. https://doi.org/10.1016/j.xpro.2023.102337