Studies on the microcytotoxicity test.  III.  Comparison of 75Se-selenomethionine with 3H-proline, Na251Cr04 and 125I-iododeoxyuridine for pre-labelling target cells in long-term cytotoxicity assays.

Brooks, C.G.
Cancer Research Laboratories, University of Nottingham, U.K.

J.Immunol.Meth. 22: 23-36, 1978.
 

Four intracellular radioisotope labels, [3H]proline, Na 2 51CrO4, [75Se]selenomethionine and [125I]iododeoxyuridine, were evaluated for use in a pre-labelling long-term microcytotoxicity assay for cell-mediated immunity. Adherent 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 Na 2 51CrO4 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 gamma-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.