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Hubrecht Organoid Technology (HUB)

3 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

Organoid-Derived Epithelial Monolayer: A Clinically Relevant In Vitro Model for Intestinal Barrier Function

Wies T. M. van Dooremalen¹, Merel Derksen¹, Jamie Lee Roos¹, Celia Higuera Barón¹, Carla S. Verissimo¹, Robert G. J. Vries¹, Sylvia F. Boj¹, Farzin Pourfarzad¹

¹Foundation Hubrecht Organoid Technology (HUB)

Here, we describe the preparation of human organoid-derived intestinal epithelial monolayers for studying intestinal barrier function, permeability, and transport. As organoids represent original epithelial tissue response to external stimuli, these models combine the advantages of expandability of cell lines and the relevance and complexity of primary tissue.

Cancer Research

Medium-Throughput Drug- and Radiotherapy Screening Assay using Patient-Derived Organoids

Marrit Putker^*1, Rosemary Millen^*2, René Overmeer^*3, Else Driehuis^2,4, Maurice M. J. M. Zandvliet⁵, Hans Clevers², Sylvia F. Boj³, Qi-Xiang Li⁶

¹Crown Bioscience Netherlands B.V., ²Oncode Institute, Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and University Medical Center Utrecht, ³Hubrecht Organoid Technology (HUB), ⁴Pathology dept, University Medical Center Utrecht, ⁵Department of Clinical Sciences - Companion Animals, Faculty of Veterinary Medicine, Utrecht University, ⁶Crown Bioscience Inc.

We describe detailed protocols to use patient-derived organoids for medium-throughput therapy sensitivity screenings. Therapies tested include chemotherapy, radiotherapy, and chemo-radiotherapy. Adenosine triphosphate levels are used as a functional readout.