Determination of thymidine in serum used for cell culture media

Thymidine concentrations in serum used for cell culture media were determined with an assay based on isotope dilution. In this assay, incorporation of (3H)-thymidine into DNA of cultured cells was measured in the presence of 5 and 20% serum as a function of the concentration of unlabeled thymidine added to the medium. In three batches of horse serum, thymidine concentrations were 0--0.17 micron, while in fetal calf serum values of 0.75--2.1 micron were obtained. Dialysis of serum resulted in a reduction of thymidine levels by factors of at least 10.