Rapid assay system for cytotoxicity tests using 14C-leucine incorporation into tumor cells.

Cell viability under various conditions of cytotoxicity test was assessed by terminal labeling of tumor cells, T24 cell line derived from urinary bladder carcinoma, with 14C-leucine. Changes of 14C-leucine incorporation into the cells were fairly proportional to those of viable cell number measured by the trypan blue exclusion method, but a definite correlation between the two measurements was not always found following cytotoxic manipulations. When the cells were labeled immediately after drug treatments, 14C-leucine incorporation usually led to fluctuated and insensitive results presumably due to disturbed metabolic activities unrelated to cell viability and failed to indicate the degree of cell damage. It was shown, however, that the cytotoxicity test was satisfactorily determined by labeling tumor cells with 14C-leucine after recovery in fresh medium for 24 hr. Cytotoxic effects of anticancer drugs with concentration- and time-dependency, hyperthermia and phytohemagglutinin-stimulated lymphocytes were demonstrated by the radioactivity incorporated into the target cells on day 2 after removal of the cytotoxic factors. The results indicate that the terminal labeling of tumor cells with 14C-leucine can be used as a rapid and reliable measure sensitive to cell viability for an in vitro assay system.