Dear all, recently i am considering if i have been doing my MTT assay wrongly as i have been using my media (with 10% FBS) for my cell viability assay. I realised that the cells were dying at high concentrations of the drug - i.e. 50 and 100 micromolar. Do you believe that by starving the cells first (with serum free media) prior to adding my drug (with 0.1% FBS) and incubating them will yield more reliable results? As in will more cells than die under these conditions? For all your kind advice, please. Thank you.
It should be totally fine to use medium + 10% FBS, unless you use different medium for your control cells.
You may try also different medium, but always keep the same conditions (except drug concentrations) in different wells. Less FBS may change the metabolism rate of cells if it is related somehow to the mechanism of action of your compound.
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