Related Experiment Video
Updated: Apr 1, 2026
Assessing Cell Viability and Death in 3D Spheroid Cultures of Cancer Cells
Published on: June 16, 2019
A 3D model for assessing regulated cell death modalities
Robin Demuynck1, Dmitri V Krysko1
1Cell Death Investigation and Therapy (CDIT) Laboratory, Anatomy and Embryology Unit, Department of Human Structure and Repair, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium; Cancer Research Institute Ghent (CRIG), Ghent, Belgium.
Abstract:
Three-dimensional (3D) models more closely represent the in vivo situation and are therefore more relevant models for drug screening. Among the more than twelve regulated cell death (RCD) modalities, immunogenic cell death (ICD) stands out for its ability to initiate efficient anti-tumor immune response. For example, ferroptosis, which can be induced via the inhibition of GPX4 leading to uncontrolled lipid peroxidation, might be immunogenic under certain conditions. It is crucial to identify which cell death modality is induced, as certain cancer types exhibit resistance to specific forms of cell death. However, a major limitation of 3D models is the lack of high-throughput assays, which often require dissociation of the 3D models, potentially leading to misinterpretation of results. Here, we describe a protocol for identifying and quantifying the induction of RCD modalities in 3D models, such as spheroids. This method eliminates the need for tumor spheroid dissociation and is compatible with other screening techniques, including confocal microscopy. This protocol enables high-throughput screening of various cell death inducers in intact 3D models, serving as a crucial first step in the identification of novel inducers and their specificity for particular ICD and RCD types.
Insights
This study introduces a high-throughput assay for screening regulated cell death (RCD) modalities, like immunogenic cell death (ICD), in intact 3D cancer models. This method avoids sample dissociation, enabling accurate drug screening and identification of novel RCD inducers.
Area of Science:
- Biomedical Sciences
- Cancer Research
- Drug Discovery
Background:
- Three-dimensional (3D) models better mimic in vivo conditions for drug screening.
- Regulated cell death (RCD) modalities, particularly immunogenic cell death (ICD), are crucial for anti-tumor immunity.
- Identifying specific RCD types is vital due to cancer's resistance to certain cell death pathways.
Purpose of the Study:
- To develop a high-throughput assay for quantifying RCD modalities in intact 3D models.
- To overcome the limitations of sample dissociation in current 3D model assays.
- To facilitate the screening of novel RCD and ICD inducers for cancer therapy.
Main Methods:
- Developed a protocol for analyzing RCD induction in 3D models (e.g., spheroids) without dissociation.
- Ensured compatibility with other screening techniques like confocal microscopy.
- Enabled high-throughput screening of cell death inducers in intact 3D models.
Main Results:
- Successfully identified and quantified RCD modalities in intact 3D models.
- The protocol eliminates the need for spheroid dissociation, preventing data misinterpretation.
- Demonstrated compatibility with standard imaging techniques for RCD analysis.
Conclusions:
- The new protocol allows for high-throughput RCD and ICD screening in 3D models.
- This method is a critical advancement for identifying novel cancer therapeutics.
- Accurate assessment of cell death in 3D models is now feasible for drug discovery.
More Related Videos
10:03Author Spotlight: Understanding Cytokine-Induced Cell Death in Intestinal Epithelial Cells Using Human Organoids
Published on: August 2, 2024
13:54Preparation of Cell-lines for Conditional Knockdown of Gene Expression and Measurement of the Knockdown Effects on E4orf4-Induced Cell Death
Published on: October 21, 2012
