Studies on the microcytotoxicity test. II. The uptake of amino acids ([3H]leucine or [75Se]methionine) but not nucleosides ([3H]thymidine or [125I]IUdR) or 51CrO24-provides a direct and quantitative measure of target cell survival in the presence of lymphoid cells.
We have studied the suitability of various commonly used radioactive materials for the direct post-labeling of adherent rat target cells in long-term cytotoxicity tests. The use of nucleosides at high concentration avoids the necessity of adding fluorodeoxyuridine to enhance nucleoside uptake by target cells, and reduces the degree of non-specific inhibition of nucleoside uptake caused by products released from effector lymphoid cells. However, when [125I]iododeoxyuridine was used for labelling, such inhibition was not completely avoided even at very high nucleoside concentration, necessitating the washing of target cells prior to labelling. Similarly, without prewashing, the uptake of 51CrO24-ions frequently failed to correlate well with the numbers of surviving target cells as assessed by cell counting. On the other hand, radiolabelled amino acids, when present at semi-saturating concentrations, were taken up quantitatively by target cells under all conditions tested. Furthermore, in comparison to [125I]iododeoxyuridine, radioactive amino acids showed little if any toxicity to target cells. The use of the gamma-emitting amino acid analogue, [75Se]selenomethionine, is particularly recommended.
D000818:Animals; D002478:Cells, Cultured; D002857:Chromium / Q000378:metabolism*; D002860:Chromium Radioisotopes; D003470:Culture Media; D003601:Cytotoxicity Tests, Immunologic; D004306:Dose-Response Relationship, Immunologic; D007065:Idoxuridine / Q000378:metabolism*; D007930:Leucine / Q000378:metabolism*; D008198:Lymph Nodes / Q000276:immunology; D008214:Lymphocytes / Q000276:immunology*; D009374:Neoplasms, Experimental / Q000276:immunology; D010087:Oxides / Q000378:metabolism; D051381:Rats; D012643:Selenium / Q000378:metabolism*; D012645:Selenomethionine / Q000378:metabolism*; D013936:Thymidine / Q000378:metabolism*
D002860:Chromium Radioisotopes; D003470:Culture Media; D010087:Oxides; D002857:Chromium; D012645:Selenomethionine; D007930:Leucine; D012643:Selenium; D007065:Idoxuridine; D013936:Thymidine