ATP-Based Luciferase Viability Assay: A Homogenous Method to Evaluate the Growth-Inhibitory Potential of Test Agents on Tumor Organoids

After 24 hours of medium change, mince the tumor organoids using a cell fragmentation and dispersion instrument, equipped with a filter holder containing a 70-micrometer mesh filter. Dilute 15 millimeters of tumor organoid suspension 10 times.

Use the cell suspension dispenser to seed 40 microliters of diluted tumor organoid suspension in a 384-well ultra-low attachment spheroid microplate.

After 24 hours, use a liquid handler to add 0.04 microliters of test agent solutions from 10 serial dilutions at final concentration ranges of 20 micromolar to 1.0 nanomolar in the wells, and incubate the plate for 6 days.

At the end of the incubation, add intracellular ATP-measuring reagent in the test wells. Use a mixer to mix the contents of the plate and incubate the plate for 10 minutes at approximately 25 degrees Celsius. Use a plate reader to measure the intracellular ATP content as luminescence.