The “Protocol for Application, Standardization and Validation of the Forskolin-Induced Swelling Assay in Cystic Fibrosis Human Colon Organoids” has been published in STAR Protocols, a Cell press journal.
The protocol, available online, describes the isolation, handling, culture of, and experiments carried on human colon stem cell organoids in the context of Cystic Fibrosis (CF). In CF, the functioning of proteins called cystic fibrosis transmembrane conductance regulator (CFTR) is impaired. 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.
This project has been supported by the Dutch Cystic Fibrosis Foundation (NCFS). These protocols have been endorsed by the European Cystic Fibrosis Society (ECFS) and by ERN-LUNG in 2019, and aim to standardise methods to ensure high reproducibility and correct interpretation of results among different laboratories.