Comparison of 3H-thymidine incorporation and CellTiter 96 aqueous colorimetric assays in cell proliferation of bovine mononuclear cells

Acta Vet Hung. 1998;46(2):191-7.


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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Cattle
  • Cells, Cultured / metabolism
  • Colorimetry / veterinary
  • Concanavalin A / pharmacology
  • Dose-Response Relationship, Drug
  • Indicators and Reagents / analysis*
  • Indicators and Reagents / metabolism
  • Lymph Nodes / chemistry
  • Lymph Nodes / cytology
  • Lymphocyte Activation / physiology*
  • Spleen / chemistry
  • Spleen / cytology
  • Tetrazolium Salts / analysis*
  • Tetrazolium Salts / metabolism
  • Thymidine / analysis*
  • Thymidine / metabolism
  • Thymus Gland / chemistry
  • Thymus Gland / cytology
  • Tritium


  • Indicators and Reagents
  • Tetrazolium Salts
  • Tritium
  • Concanavalin A
  • Thymidine