JoVE Encyclopedia of Experiments
Cancer Research
0 views • 5:39 min • April 30th, 2023
- Pancreatic cancer cells interact with cancer-associated fibroblasts in the tumor microenvironment or TME. These interactions modulate the metabolic activity of the cancer cells and affect cancer progression.
Spheroids are the cellular aggregates that recapitulate the TME in vitro. Therefore, we can use these spheroids to test the changes in tumor's metabolic functions by extracellular flux assay. This assay measures the extracellular concentration of oxygen and protons to determine aerobic and glycolytic metabolism, respectively.
Begin by taking appropriately sized 3D spheroids made by co-culturing pancreatic cancer cells with pancreatic fibroblasts. Now, transfer the spheroids into a flux analysis plate and cover it with a sensor cartridge containing test drugs. Run the assay in an extracellular flux analyzer.
Flux analyzer pneumatically injects test drugs from the sensor cartridge to the analysis plate at specific time points. In response to the test drugs, cells release protons and oxygen into the plate micro-chamber and the fluorophores present in the sensor probe measure these levels. Finally, using appropriate software, analyze the functionality of the spheroids.
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