Protocol for Application, Standardization and Validation of the Forskolin-Induced Swelling Assay in Cystic Fibrosis Human Colon Organoids
Publication date
2020-06-19
Authors
Vonk, Annelotte M
Van Mourik, Peter
Ramalho, Anabela S
Silva, Iris A L
Statia, Marvin
Kruisselbrink, E
Suen, Sylvia W F
Dekkers, Johanna F
Vleggaar, Frank P
Houwen, R. H J
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Advisors
Supervisors
Document Type
Article
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cc_by_nc_nd
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).
Keywords
General Biochemistry,Genetics and Molecular Biology, General Neuroscience, General Immunology and Microbiology, Journal Article
Citation
Vonk, A M, van Mourik, P, Ramalho, A S, Silva, I A L, Statia, M, Kruisselbrink, E, Suen, S W F, Dekkers, J F, Vleggaar, F P, Houwen, R H J, Mullenders, J, Boj, S F, Vries, R, Amaral, M D, de Boeck, K, van der Ent, C K & Beekman, J M 2020, 'Protocol for Application, Standardization and Validation of the Forskolin-Induced Swelling Assay in Cystic Fibrosis Human Colon Organoids', STAR protocols, vol. 1, no. 1, 100019, pp. 1-31. https://doi.org/10.1016/j.xpro.2020.100019