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Protocol for generating airway organoids from 2D air liquid interface-differentiated nasal epithelia for use in a functional CFTR assay
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 describ...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10511861/ https://www.ncbi.nlm.nih.gov/pubmed/37314920 http://dx.doi.org/10.1016/j.xpro.2023.102337 |
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author | Rodenburg, Lisa W. van der Windt, Isabelle S. Dreyer, Henriette H.M. Smits, Shannon M.A. den Hertog - Oosterhoff, Loes A. Aarts, Ellen M. Beekman, Jeffrey M. Amatngalim, Gimano D. |
author_facet | Rodenburg, Lisa W. van der Windt, Isabelle S. Dreyer, Henriette H.M. Smits, Shannon M.A. den Hertog - Oosterhoff, Loes A. Aarts, Ellen M. Beekman, Jeffrey M. Amatngalim, Gimano D. |
author_sort | Rodenburg, Lisa W. |
collection | PubMed |
description | 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) |
format | Online Article Text |
id | pubmed-10511861 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-105118612023-09-22 Protocol for generating airway organoids from 2D air liquid interface-differentiated nasal epithelia for use in a functional CFTR assay Rodenburg, Lisa W. van der Windt, Isabelle S. Dreyer, Henriette H.M. Smits, Shannon M.A. den Hertog - Oosterhoff, Loes A. Aarts, Ellen M. Beekman, Jeffrey M. Amatngalim, Gimano D. STAR Protoc Protocol 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) Elsevier 2023-06-12 /pmc/articles/PMC10511861/ /pubmed/37314920 http://dx.doi.org/10.1016/j.xpro.2023.102337 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Protocol Rodenburg, Lisa W. van der Windt, Isabelle S. Dreyer, Henriette H.M. Smits, Shannon M.A. den Hertog - Oosterhoff, Loes A. Aarts, Ellen M. Beekman, Jeffrey M. Amatngalim, Gimano D. Protocol for generating airway organoids from 2D air liquid interface-differentiated nasal epithelia for use in a functional CFTR assay |
title | Protocol for generating airway organoids from 2D air liquid interface-differentiated nasal epithelia for use in a functional CFTR assay |
title_full | Protocol for generating airway organoids from 2D air liquid interface-differentiated nasal epithelia for use in a functional CFTR assay |
title_fullStr | Protocol for generating airway organoids from 2D air liquid interface-differentiated nasal epithelia for use in a functional CFTR assay |
title_full_unstemmed | Protocol for generating airway organoids from 2D air liquid interface-differentiated nasal epithelia for use in a functional CFTR assay |
title_short | Protocol for generating airway organoids from 2D air liquid interface-differentiated nasal epithelia for use in a functional CFTR assay |
title_sort | protocol for generating airway organoids from 2d air liquid interface-differentiated nasal epithelia for use in a functional cftr assay |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10511861/ https://www.ncbi.nlm.nih.gov/pubmed/37314920 http://dx.doi.org/10.1016/j.xpro.2023.102337 |
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