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Article

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

by
Annelies Van Hemelryk
1,
Sigrun Erkens-Schulze
1,
Lifani Lim
1,
Corrina M. A. de Ridder
1,
Debra C. Stuurman
1,
Guido W. Jenster
1,
Martin E. van Royen
2,† and
Wytske M. van Weerden
1,*,†
1
Department of Urology, Erasmus University Medical Center, Dr. Molewaterplein 40, 3015 GD Rotterdam, The Netherlands
2
Department of Pathology, Erasmus University Medical Center, Dr. Molewaterplein 40, 3015 GD Rotterdam, The Netherlands
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Cells 2023, 12(10), 1377; https://doi.org/10.3390/cells12101377
Submission received: 21 April 2023 / Revised: 8 May 2023 / Accepted: 11 May 2023 / Published: 12 May 2023
(This article belongs to the Collection Advances in 3D Cell Culture)
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Abstract

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.
Keywords: organoid; drug testing; viability assays; live-cell confocal microscopy; high-content screening; prostate cancer; patient-derived xenograft organoid; drug testing; viability assays; live-cell confocal microscopy; high-content screening; prostate cancer; patient-derived xenograft

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MDPI and ACS Style

Van Hemelryk, A.; Erkens-Schulze, S.; Lim, L.; de Ridder, C.M.A.; Stuurman, D.C.; Jenster, G.W.; van Royen, M.E.; van Weerden, W.M. Viability Analysis and High-Content Live-Cell Imaging for Drug Testing in Prostate Cancer Xenograft-Derived Organoids. Cells 2023, 12, 1377. https://doi.org/10.3390/cells12101377

AMA Style

Van Hemelryk A, Erkens-Schulze S, Lim L, de Ridder CMA, Stuurman DC, Jenster GW, van Royen ME, van Weerden WM. Viability Analysis and High-Content Live-Cell Imaging for Drug Testing in Prostate Cancer Xenograft-Derived Organoids. Cells. 2023; 12(10):1377. https://doi.org/10.3390/cells12101377

Chicago/Turabian Style

Van Hemelryk, Annelies, Sigrun Erkens-Schulze, Lifani Lim, Corrina M. A. de Ridder, Debra C. Stuurman, Guido W. Jenster, Martin E. van Royen, and Wytske M. van Weerden. 2023. "Viability Analysis and High-Content Live-Cell Imaging for Drug Testing in Prostate Cancer Xenograft-Derived Organoids" Cells 12, no. 10: 1377. https://doi.org/10.3390/cells12101377

APA Style

Van Hemelryk, A., Erkens-Schulze, S., Lim, L., de Ridder, C. M. A., Stuurman, D. C., Jenster, G. W., van Royen, M. E., & van Weerden, W. M. (2023). Viability Analysis and High-Content Live-Cell Imaging for Drug Testing in Prostate Cancer Xenograft-Derived Organoids. Cells, 12(10), 1377. https://doi.org/10.3390/cells12101377

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