Comparison of MTT colorimetric assay and tritiated thymidine uptake for lymphocyte proliferation assays using chicken splenocytes.
The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) colorimetric assay was compared with the conventional tritiated thymidine deoxyriboside (3H-TdR) incorporation for assay of lymphocyte blastogenesis using mononuclear cells isolated from the spleens of specific-pathogen-free chickens. The study was undertaken in an effort to simplify methods for assessing avian lymphocyte proliferation, specifically for evaluating response to mitogens or for indirect measurement of T-cell growth factors. The results from stimulated cells in both assay methods were significantly different from results from the control cells, and the MTT assay results regressed in a significant linear manner on counts from 3H-TdR incorporation. On this basis, the MTT assay is a valid test for evaluation of lymphocyte proliferation of chicken splenocytes.
Bounous
D I
DI
Department of Pathology, College of Veterinary Medicine, University of Georgia, Athens 30602.
Campagnoli
R P
RP
Brown
J
J
D000818:Animals; D002478:Cells, Cultured; D002645:Chickens / Q000276:immunology*; D003124:Colorimetry / Q000379:methods / Q000662:veterinary*; D004396:Coloring Agents; D007958:Leukocyte Count / Q000662:veterinary; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000378:metabolism; D013154:Spleen / Q000166:cytology; D013223:Statistics as Topic; D013778:Tetrazolium Salts; D013844:Thiazoles; D013936:Thymidine; D014316:Tritium
D004396:Coloring Agents; D013778:Tetrazolium Salts; D013844:Thiazoles; D014316:Tritium; C022616:thiazolyl blue; D013936:Thymidine