Is it plausible that cells exposed to an oxidative stressor would display increased metabolism (as measured through MTT assays)?
Hello Fiona Burmeister Mccracken
Changes in the activity of dozens of mitochondrial and non-mitochondrial oxidoreductases, cellular metabolic and energy perturbations, and oxidative stress may significantly impact the MTT assay readout.
A level of glucose, its uptake rate and rate of glycolysis, a level of lactate, pyruvate, and NADH/NADPH is known to influence MTT reduction. MTT assay do not actually measure the number of viable cells or their growth but rather an integrated set of enzyme activities that are related in various ways to cell metabolism. Also, the factors changing a rate of endo-/exocytosis and intracellular trafficking of MTT-formazan may affect MTT reduction.
Therefore, one should be concerned by the possibility that a degree of viability change judged from the observed changes in MTT reduction between control and experimental cells can be over/underestimated due to the influence of experimental manipulations on the parameters mentioned above rather than due to a real viability decrease.
So, to avoid result misinterpretation, supplementation of the tetrazolium salt-based assays with other non-metabolic assays is recommended.
You may want to refer to the article attached below for more information.
https://www.sciencedirect.com/science/article/abs/pii/S037811191500952X
Best.
Hello,
I totaly agree with Malcolm. the MTT test suffers from numerous interferences, particularly in the case of modified oxido-reducing ratios.
Take the problem in another direction. Instead of counting living cells by MTT, you could count cells that have died, by measuring in the medium markers of cell lysis, such as LDH activity, or released DNA.
Good luck. All the best.
Marc
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