Studies on the microcytotoxicity test. 3

Four intracellular radioisotope labels, [3H]proline, Na251CrO4, [75Se]selenomethionine and [125I]iododeoxyuridine, were evaluated for use in a pre-labelling long-term microcytotoxicity assay for cell-mediated immunity. Adherent rat tumour cells established in tissue culture were used as targets and the basic variables studied were labelling efficiency, toxicity and spontaneous release rates. [125I]Iododeoxyuridine was found unsuitable on account of its high toxicity and correspondingly high spontaneous release rate, and Na251CrO4 for its toxicity and low labelling efficiency. Of the two other radiolabels, [75Se]selenomethionine had the advantage over [3H]proline of higher labelling efficiency (especially in Ham's F10 medium), lower toxicity, and being a γ-emitter. Furthermore, released 75Se was shown to be non-reutilisable and its retention by target cells provided an accurate measure of cell survival in an alloimmune system. Methods of calculating the results of pre-labelling cytotoxicity tests based on the total radioactivity in target cells at the beginning of the assay were found to be invalid. (Auth.)