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Updated: Jul 15, 2026
Assessing Cell Viability and Death in 3D Spheroid Cultures of Cancer Cells
Published on: June 16, 2019
High throughput assessment of viability in 2D and 3D cell culture
Flora Doffe1, Marion Leduc1, Xiaolian Deng1
1Institut Universitaire de France, Université Paris Cité, Sorbonne Université, Inserm, Centre de Recherche des Cordeliers, Equipe labellisée par la Ligue contre le cancer, Paris, France; Institut Gustave Roussy, Université Paris-Saclay, INSERM US23/CNRS UAR 3655, Metabolomics and Cell Biology Platforms, Villejuif, France.
Abstract:
High-throughput screening (HTS) plays an essential role in oncological drug discovery by facilitating the identification of novel anticancer agents through the evaluation of cytotoxic responses in malignant cells. Advances in automation, imaging technologies, and labware design have significantly enhanced the feasibility and scalability of HTS in cell 2D as well as 3D cell culture systems. However, accessible and systematic dose-finding approaches remain a major unmet need, often constraining the interpretability and reproducibility of screening outcomes. In this study, we present robust high-throughput screening assays that employ standard fluorescent staining and ATP-limited conversion methods to quantify a range of cellular parameters associated with cell health and death. This strategy enables multiparametric evaluation of compound effects, enhancing the ability to distinguish between general cytotoxicity and targeted therapeutic efficacy. Our method is compatible with existing HTS workflows and provides a scalable solution for early-stage drug discovery efforts.
Insights
This study introduces robust high-throughput screening (HTS) assays for cancer drug discovery. These methods improve the analysis of cell health and death, aiding in the identification of effective anticancer agents.
Area of Science:
- Oncology
- Pharmacology
- Biotechnology
Background:
- High-throughput screening (HTS) is crucial for identifying anticancer drugs by assessing cytotoxicity in cancer cells.
- Advancements in automation and imaging have improved HTS scalability in 2D and 3D cell cultures.
- A need exists for systematic dose-finding methods to enhance HTS interpretability and reproducibility.
Purpose of the Study:
- To develop robust HTS assays for multiparametric evaluation of compound effects in cancer drug discovery.
- To improve the distinction between general cytotoxicity and targeted therapeutic efficacy.
- To provide a scalable solution compatible with existing HTS workflows.
Main Methods:
- Utilized standard fluorescent staining techniques.
- Employed ATP-limited conversion methods for quantifying cellular parameters.
- Developed assays to measure cell health and death indicators.
Main Results:
- Established reliable HTS assays for evaluating cellular responses to potential drug compounds.
- Enabled multiparametric assessment of compound-induced cytotoxicity.
- Demonstrated compatibility with existing HTS infrastructure.
Conclusions:
- The developed HTS assays offer a scalable and robust approach for early-stage oncological drug discovery.
- The method enhances the interpretability and reproducibility of screening outcomes.
- This strategy aids in identifying novel anticancer agents with improved precision.
