Protocol for Application, Standardization and Validation of the Forskolin-Induced Swelling Assay in Cystic Fibrosis Human Colon Organoids

Annelotte M Vonk, Peter van Mourik, Anabela S Ramalho, Iris A L Silva, Marvin Statia, Evelien Kruisselbrink, Sylvia W F Suen, Johanna F Dekkers, Frank P Vleggaar, Roderick H J Houwen, Jasper Mullenders, Sylvia F Boj, Robert Vries, Margarida D Amaral, Kris de Boeck, Cornelis K van der Ent, Jeffrey M Beekman

Research output: Contribution to journalArticleAcademicpeer-review

16 Citations (Scopus)
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Abstract

This protocol describes the isolation, handling, culture of, and experiments with human colon stem cell organoids in the context of cystic fibrosis (CF). In human colon organoids, the function of cystic fibrosis transmembrane conductance regulator (CFTR) protein and its rescue by CFTR modulators can be quantified using the forskolin-induced swelling assay. Implementation procedures and validation experiments are described for six CF human colon organoid lines, and representative CFTR genotypes are tested for basal CFTR function and response to CFTR-modulating drugs. For complete details on the use and execution of this protocol, please refer to Dekkers et al (2016) and Berkers and van Mourik (2019).

Original languageEnglish
Article number100019
Pages (from-to)1-31
Number of pages31
JournalSTAR protocols
Volume1
Issue number1
DOIs
Publication statusPublished - 19 Jun 2020

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