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Updated: Feb 22, 2026
Assessing Cell Viability and Death in 3D Spheroid Cultures of Cancer Cells
Published on: June 16, 2019
Real-Time Apoptosis and Viability High-Throughput Screening of 3D Multicellular Tumor Spheroids Using the Celigo
Sarah Kessel1, Scott Cribbes1, Surekha Bonasu1
11 Department of Technology R&D, Nexcelom Bioscience LLC, Lawrence, MA, USA.
Abstract:
Three-dimensional tumor spheroid models have been increasingly used to investigate and characterize cancer drug compounds. Previously, the Celigo image cytometer has demonstrated its utility in a high-throughput screening manner for evaluating potential drug candidates in a 3D multicellular tumor spheroid (MCTS) primary screen. In addition, we have developed real-time kinetic caspase 3/7 apoptosis and propidium iodide viability 3D MCTS assays, both of which can be used in a secondary screen to better characterize the hit compounds. In this work, we monitored the kinetic apoptotic and cytotoxic effects of 14 compounds in 3D MCTS produced from the glioblastoma cell line U87MG in 384-well plates for 9 days. The kinetic results allowed the categorization of the effects from 14 drug compounds into early and late cytotoxic, apoptotic, cytostatic, and no effects. The real-time apoptosis and viability screening method can serve as an improved secondary screen to better understand the mechanism of action of these potential drug candidates identified from the primary screen, allowing one to identify a more qualified drug candidate and streamline the drug discovery process of research and development.
Insights
This study introduces a real-time assay for evaluating cancer drug compounds in 3D tumor models. The method categorizes drug effects, improving the selection of qualified drug candidates for streamlined drug discovery.
Area of Science:
- Oncology
- Pharmacology
- Biotechnology
Background:
- Three-dimensional tumor spheroid models are crucial for cancer drug discovery.
- The Celigo image cytometer facilitates high-throughput screening of 3D multicellular tumor spheroids (MCTS).
- Real-time kinetic assays for apoptosis and viability enhance secondary screening of drug candidates.
Purpose of the Study:
- To develop and validate a real-time kinetic assay for characterizing drug effects in 3D MCTS.
- To categorize the apoptotic and cytotoxic effects of 14 compounds on glioblastoma U87MG MCTS.
- To improve the secondary screening process for identifying qualified drug candidates and streamlining R&D.
Main Methods:
- Utilized 3D MCTS derived from the U87MG glioblastoma cell line in 384-well plates.
- Monitored kinetic apoptotic (caspase 3/7) and viability (propidium iodide) effects over 9 days.
- Employed real-time screening to categorize compound effects as early/late cytotoxic, apoptotic, cytostatic, or none.
Main Results:
- Successfully categorized the kinetic effects of 14 compounds on U87MG 3D MCTS.
- Distinguished between early and late cytotoxic, apoptotic, and cytostatic responses.
- Demonstrated the utility of real-time kinetic assays for detailed compound characterization.
Conclusions:
- The real-time apoptosis and viability screening method serves as an improved secondary screen for drug discovery.
- This approach enhances the understanding of drug mechanisms of action.
- Streamlines the identification of qualified drug candidates, optimizing the research and development process.
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