Thymidine resistance in Chinese hamster V79 cells in vitro

Thymidine resistance in V79 Chinese hamster cells has been investigated. Phenotypically stable variant resistant lines occurred at a high frequency, and the mutation rate (2.67 x 10(-3) per cell per generation) to 400 micrograms/ml thymidine resistance as measured by the standard Luria--Delbrück fluctuation analysis was extremely high. Populations of cells maintained for extended periods in F-10 medium spontaneously increased in resistance, possibly as a result of selective pressures due to the thymidine present in F-10 medium since this change was not observed in Dulbecco's medium. The degree of resistance for a given variant was correlated with the amount of thymidine employed in its selection. Metabolic cooperation, resulting in the suppression of the resistant phenotype, was demonstrated in artificial mixtures of sensitive and resistant clonal lines. Clones isolated in high levels of thymidine possessed lowered uptake of [3H]thymidine and the depression in uptake was related to the level of resistance of the particular clone. Although thymidine kinase specific activity levels were slightly depressed in variant cell lines, growth rate and uridine uptake were unaffected. We conclude that thymidine resistance is due to a genetically controlled depression of external thymidine uptake.