Comparison of 3H-thymidine incorporation and CellTiter 96 aqueous colorimetric assays in cell proliferation of bovine mononuclear cells.
A rapid colorimetric non-radioactive assay for the determination of bovine mitogen-induced lymphocyte proliferation in cell culture was evaluated using a novel tetrazolium compound (MTS) and an electron coupling reagent (PMS) provided in the CellTiter 96 kit (Promega). The results of the new method were compared with those of the 3H-thymidine incorporation assay using parallels obtained from the same lymphocyte population. The concentrations used in the cell suspension of primary cultured lymphocytes resulted in a significant signal/background ratio when cells were prepared from peripheral blood, spleen or mesenteric lymph nodes. The same concentrations of thymocytes resulted in a weak signal even for the highest concentrations of mitogen. A good correlation was demonstrated between the results of the two methods. The non-radioactive method performed well in assays in bovine mononuclear cells derived from prolactin-treated or -untreated calves, showing a 50% lower responsiveness to mitogenic stimulation in prolactin-deprived animals.
Zolnai
A
A
Department of Physiology and Biochemistry, University of Veterinary Science, Budapest, Hungary.
Tóth
E B
EB
Wilson
R A
RA
Frenyó
V L
VL
D000818:Animals; D000831:Animals, Newborn; D002417:Cattle; D002478:Cells, Cultured / Q000378:metabolism; D003124:Colorimetry / Q000662:veterinary; D003208:Concanavalin A / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D007202:Indicators and Reagents / Q000032:analysis* / Q000378:metabolism; D008198:Lymph Nodes / Q000737:chemistry / Q000166:cytology; D008213:Lymphocyte Activation / Q000502:physiology*; D013154:Spleen / Q000737:chemistry / Q000166:cytology; D013778:Tetrazolium Salts / Q000032:analysis* / Q000378:metabolism; D013936:Thymidine / Q000032:analysis* / Q000378:metabolism; D013950:Thymus Gland / Q000737:chemistry / Q000166:cytology; D014316:Tritium
D007202:Indicators and Reagents; D013778:Tetrazolium Salts; D014316:Tritium; D003208:Concanavalin A; D013936:Thymidine