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Forskolin-induced Swelling in Intestinal Organoids: An In Vitro Assay for Assessing Drug Response in Cystic Fibrosis Patients
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Forskolin-induced Swelling in Intestinal Organoids: An In Vitro Assay for Assessing Drug Response in Cystic Fibrosis Patients

Forskolin-induced Swelling in Intestinal Organoids: An In Vitro Assay for Assessing Drug Response in Cystic Fibrosis Patients

DOI:
10.3791/55159-v

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07:04 min

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February 11, 2017

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, , , , , , ,
1Foundation Hubrecht Organoid Technology, 2Department of Pediatric Pulmonology, Regenerative Medicine Centre Utrecht, Wilhelmina Children’s Hospital,University Medical Centre Utrecht, 3Department of Stem Cells and Cancer,Walter and Eliza Hall Institute of Medical Research, 4Hubrecht Institute for Developmental Biology and Stem Cell Research,University Medical Centre Utrecht

Chapters

  • 00:05Title
  • 01:06Colon Organoid Passaging
  • 03:56Forskolin-induced Swelling Assay (FIS)
  • 05:29Results: Representative FIS Quantification
  • 06:14Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes an assay for measuring CFTR function and CFTR modulator responses in cultured tissue from subjects with cystic fibrosis (CF). Biopsy-derived intestinal organoids swell in a cAMP-driven fashion, a response that is defective (or strongly reduced) in CF organoids and can be restored by exposure to CFTR modulators.

Tags

Keywords: Forskolin-induced SwellingIntestinal OrganoidsCystic FibrosisCFTR FunctionCFTR Modulating TreatmentsFIS AssayOrthotherapyDrug Efficacy TestingOrganoid CultureBasement Membrane MatrixBasal MediumCentrifugationPipettingSample Preparation

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