Accedi

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

3 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Organoids as Model for Infectious Diseases: Culture of Human and Murine Stomach Organoids and Microinjection of Helicobacter Pylori

Sina Bartfeld¹, Hans Clevers¹

¹Hubrecht Institute for Developmental Biology and Stem Cell Research, University Medical Centre Utrecht

Stem cell derived cultures harbor tremendous potential to model infectious diseases. Here, the culture of mouse and human gastric organoids derived from adult stem cells is described. The organoids are microinjected with the gastric pathogen Helicobacter pylori.

Medicine

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.

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.