A comparison between two methods for measuring tumor necrosis factor in biological fluids

Abstract

The current study was undertaken to compare two methods for the efficiency of measuring tumor necrosis factor (TNF-α) in biological fluids, which is species undependent, reliable, sensitive, simple and not expensive. We have compared the MTT tetrazolium cytotoxic assay [1,2] and the³H-thymidine (³H-TdR) incorporation cytostatic assay for measuring the anti-tumor activity of human recombinant TNF-α, of human colonic tissue and of supernatants ofin vitro stimulated human and rat peritoneal macrophages. Two target cell-lines, namely murine myelomonocytic leukaemia WEHI-164- and L-929-transformed murine fibroblast cell-lines, were used in the MTT assay. The L-929 line was also used in the³H-TdR assay. WEHI-164 was more sensitive than the L-929 cell-line in the MTT cytotoxic assay. Furthermore, the MTT assay was more sensitive to TNF-α than the³H-TdR assay. Both methods can be used for the detection of anti-tumor activity in biological fluids but the MTT cytotoxic method has the advantage of being more sensitive and more simple.