Cancer Genomics Center (CGC)
2 ARTICLES PUBLISHED IN JoVE
JoVE Core
Forskolin-induced Swelling in Intestinal Organoids: An In Vitro Assay for Assessing Drug Response in Cystic Fibrosis Patients
Sylvia F. Boj 1, Annelotte M. Vonk 2, Marvin Statia 1, Jinyi Su 1, Johanna F. Dekkers 3, Robert R. G. Vries 1, Jeffrey M. Beekman 2, Hans Clevers 1,4
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
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.
Biology
Single-Cell Resolution Three-Dimensional Imaging of Intact Organoids
Ravian L. van Ineveld *1,2,3, Hendrikus C.R. Ariese *1,2,3, Ellen J. Wehrens 1,2,3, Johanna F. Dekkers *1,2,3,4, Anne C. Rios *1,2,3
1Princess Máxima Center for Pediatric Oncology, 2Department of Cancer Research, Oncode Institute, Hubrecht Institute-KNAW Utrecht, 3Cancer Genomics Center (CGC), 4Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and University Medical Center (UMC) Utrecht
The entire 3D structure and cellular content of organoids, as well as their phenotypic resemblance to the original tissue can be captured using the single-cell resolution 3D imaging protocol described here. This protocol can be applied to a wide range of organoids varying in origin, size and shape.
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