Entrar

Royal Netherlands Academy of Arts and Sciences (KNAW) and University Medical Center (UMC) Utrecht

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. Boj¹, Annelotte M. Vonk², Marvin Statia¹, Jinyi Su¹, Johanna F. Dekkers³, Robert R. G. Vries¹, Jeffrey M. Beekman², 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.