Viability Analysis and High-Content Live-Cell Imaging for Drug Testing in Prostate Cancer Xenograft-Derived Organoids

Research output: Contribution to journalArticleAcademicpeer-review

4 Citations (Scopus)
16 Downloads (Pure)


Tumor organoids have been pushed forward as advanced model systems for in vitro oncology drug testing, with the eventual goal to direct personalized cancer treatments. However, drug testing efforts suffer from a large variation in experimental conditions for organoid culturing and organoid treatment. Moreover, most drug tests are restricted to whole-well viability as the sole read-out, thereby losing important information about key biological aspects that might be impacted due to the use of administered drugs. These bulk read-outs also discard potential inter-organoid heterogeneity in drug responses. To tackle these issues, we developed a systematic approach for processing organoids from prostate cancer (PCa) patient-derived xenografts (PDXs) for viability-based drug testing and identified essential conditions and quality checks for consistent results. In addition, we generated an imaging-based drug testing procedure using high-content fluorescence microscopy in living PCa organoids to detect various modalities of cell death. Individual organoids and cell nuclei in organoids were segmented and quantified using a dye combination of Hoechst 33342, propidium iodide and Caspase 3/7 Green, allowing the identification of cytostatic and cytotoxic treatment effects. Our procedures provide important insights into the mechanistic actions of tested drugs. Moreover, these methods can be adapted for tumor organoids originating from other cancer types to increase organoid-based drug test validity, and ultimately, accelerate clinical implementation.

Original languageEnglish
Article number1377
Issue number10
Publication statusPublished - 12 May 2023

Bibliographical note

Funding Information:
This work was supported by the Translational Research Network for Prostate Cancer (TransPot) and funded by the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement No 721746 (W.M.v.W.). Financial support was also provided by the Movember Foundation, as part of the Global Action Plan 1 (GAP1) PDX project (W.M.v.W.). The Opera Phenix High Content Screening System was funded by the Daniel den Hoed Foundation.

Publisher Copyright:
© 2023 by the authors.


Dive into the research topics of 'Viability Analysis and High-Content Live-Cell Imaging for Drug Testing in Prostate Cancer Xenograft-Derived Organoids'. Together they form a unique fingerprint.

Cite this