A Rapid Cytotoxicity Assay of Metabolic Inhibitors

Abstract

H₂0₂ was produced upon the addition of menadione to mammalian cell culture. Chemiluminescent assay (CL assay) of menadione-catalyzed H₂0₂ production by viable cells was useful for the rapid detection of the cytotoxic inhibitors because the assay require only 10 min and detected the immidiate cytotoxic effect occurring a few minutes after the addition of inhibitors to cell culture. This study demonstrated that the chemiluminescent assay was superior to NR (neutral red) inclusion assay and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reduction assay on the basis of the sensitivity and rapidicity for the detection of cytotoxic effects of antibiotics and metabolic inhibitors such as DNA synthesis inhibitors, protein synthesis inhibitors, ATPase inhibitors and ionophorous antibiotics. Furthermore, the trypsinized cells were found to be more sensitive to the above inhibitors than the adherent cells with the chemiluminescent assay.