In Vitro Toxicity in Primary Adult Rat Hepatocyte Cultures
Stout, Cheryl L.
MetadataShow full item record
Primary adult rat hepatocyte monolayer cultures were used to study the toxic effects of methylmethanesulfonate (MMS) and lysolecithin. Three methods were used to evaluate the possible cytotoxic effects of the two drugs: release of two intracellular enzymes, lactate dehydrogenase (LDH) and serum glutamic oxaloacetate transaminase (SGOT); trypan blue exclusion; and scanning electron microscopy. Cultures were treated with 50, 100, and 200 µg/ml MMS and 0.5 ml aliquots of medium were removed after 2 and 6 hours of incubation for the enzyme analysis. Cultures were treated with lysolecithin at concentrations at 1.0, 15, 50, and 200 µg/ml and 0.5 ml aliquots of medium were withdrawn at 1, 3, 6, and 24 hours after treatment. Trypan blue exclusion and scanning electron microscopy were performed after all aliquots of medium had been removed for enzyme assay. A time- and dose-dependent cytotoxic effect of both drugs was demonstrated. The results indicate that a combination of methods to evaluate cytotoxicity is advantageous and that the cell culture system is an excellent method for screening drugs for toxicity.