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ATP-Based Luciferase Viability Assay: A Homogenous Method to Evaluate the Growth-Inhibitory Potential of Test Agents on Tumor Organoids
Overview
In this video, we demonstrate a rapid, single-addition assay to assess cell viability in patient-derived organoids upon treatment with a test agent using intracellular ATP levels as a marker for cellular metabolic activity. The intracellular ATP levels are measured using the luciferase/luciferin reaction, where the emitted light intensity indicates the cellular viability and, in turn, the test agent's growth-inhibitory potency.
Protocol
1. Growth Inhibition High-Throughput Screening (HTS)
NOTE: The growth inhibitory activity of anticancer agents against patient-derived tumor organoids (PDOs) is evaluated by measuring the intracellular ATP content, as shown in Figure 1. This step is performed using a commercially available cell viability assay kit (see Table of Materials).
- On day 0, culture the PDOs (e.g., RLUN007) in flasks until adequate numbers of cell clusters are available for the assay. One day before seeding, transfer the PDO suspension from a 75 cm2 flask to a 15 mL tube and centrifuge at 200 ....
Representative Results
Figure 1: Summary of the protocol used to create a high-throughput assay system using 384-well microplates.
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Source: Higa, A. et al. High-Throughput In Vitro Assay using Patient-Derived Tumor Organoids. J. Vis. Exp. (2021)
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