A comparison between two methods for measuring tumor necrosis factor in biological fluids.
The current study was undertaken to compare two methods for the efficiency of measuring tumor necrosis factor (TNF-alpha) in biological fluids, which is species undependent, reliable, sensitive, simple and not expensive. We have compared the MTT tetrazolium cytotoxic assay [1,2] and the 3H-thymidine (3H-TdR) incorporation cytostatic assay for measuring the anti-tumor activity of human recombinant TNF-alpha, of human colonic tissue and of supernatants of in 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 the 3H-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-alpha than the 3H-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.
Garrelds
I M
IM
Dept. of Pharmacology, Erasmus University, Rotterdam, The Netherlands.
Zijlstra
F J
FJ
Tak
C J
CJ
Bonta
I L
IL
Beckmann
I
I
Ben-Efraim
S
S
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D007739:L Cells; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D051381:Rats; D011994:Recombinant Proteins / Q000032:analysis; D013778:Tetrazolium Salts / Q000633:toxicity; D013844:Thiazoles / Q000633:toxicity; D013936:Thymidine / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis* / Q000633:toxicity
D011994:Recombinant Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D014409:Tumor Necrosis Factor-alpha; C022616:thiazolyl blue; D013936:Thymidine