I was doing MTT assay, and in the last day before adding the MTT; the cells looked good (or so I think), but after aspiration of the drug and addition of MTT dye I noticed that most cells seemed dead; including control cells. I would appreciate any ideas or suggestions on what might be the reason for this
https://www.ncbi.nlm.nih.gov/books/NBK144065/
Toxicity of the MTT compound is likely related to the concentration added to cells. Optimizing the concentration may result in lower toxicity. Given the cytotoxic nature of MTT, the assay method must be considered as an endpoint assay. A recent report speculated that formazan crystals contribute to harming cells by puncturing membranes during exocytosis
https://www.ncbi.nlm.nih.gov/books/NBK144065/
Toxicity of the MTT compound is likely related to the concentration added to cells. Optimizing the concentration may result in lower toxicity. Given the cytotoxic nature of MTT, the assay method must be considered as an endpoint assay. A recent report speculated that formazan crystals contribute to harming cells by puncturing membranes during exocytosis
https://pubmed.ncbi.nlm.nih.gov/22401948/
Article Exocytosis of MTT formazan could exacerbate cell injury
Although it is widely used, the MTT reagent exhibits cytotoxic effects, and adding the reagent to estimate cell viability may actually be damaging or even killing cells during the course of an experiment
https://worldwide.promega.com/resources/pubhub/is-your-mtt-assay-really-the-best-choice/
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