[HTML][HTML] Drugs with anti-oxidant properties can interfere with cell viability measurements by assays that rely on the reducing property of viable cells

N Shenoy, M Stenson, J Lawson, J Abeykoon… - Laboratory …, 2017 - nature.com
N Shenoy, M Stenson, J Lawson, J Abeykoon, M Patnaik, X Wu, T Witzig
Laboratory Investigation, 2017nature.com
Cell viability assays such as Cell Titer Blue and Alamar Blue rely on the reducing property of
viable cells to reduce the reagent dye to a product which gives a fluorescent signal. The
current manufacture-recommended protocols do not take into account the possibility of the
reagent substrate being reduced directly to the fluorescent product by drugs with an anti-
oxidant property. After suspecting spurious results while determining the cytotoxic potential
of a drug of interest (DOI) with known anti-oxidant property against a renal cell cancer (RCC) …
Abstract
Cell viability assays such as Cell Titer Blue and Alamar Blue rely on the reducing property of viable cells to reduce the reagent dye to a product which gives a fluorescent signal. The current manufacture-recommended protocols do not take into account the possibility of the reagent substrate being reduced directly to the fluorescent product by drugs with an anti-oxidant property. After suspecting spurious results while determining the cytotoxic potential of a drug of interest (DOI) with known anti-oxidant property against a renal cell cancer (RCC) cell line, we aimed to establish that drugs with anti-oxidant property can indeed cause false-negative results with the current protocols of these assays by direct reduction of the reagent substrate. We also aimed to counter the same with a simple modification added to the protocol. Through our experiments, we conclusively demonstrate that drugs with anti-oxidant properties can indeed interfere with cell viability measurements by assays that rely on the reducing property of viable cells. A simple modification in the protocol, as elaborated in the manuscript, can prevent spurious results with these otherwise convenient assays.
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