The use of ATP bioluminescence as a measure of cell proliferation and cytotoxicity

J Immunol Methods. 1993 Mar 15;160(1):81-8. doi: 10.1016/0022-1759(93)90011-u.


Adenosine triphosphate (ATP) bioluminescence was used to determine whether there was a linear relationship between cultured cell number and measured luminescence using the luciferin-luciferase reaction. In all the cells tested including peripheral blood mononuclear cells (MNC), MOLT-4, HL-60, TF-1, NFS-60 and L-929 cell lines there was a significant correlation as determined by Spearman's rank correlation coefficient (p > 0.00001). These observations were then used to determine whether ATP bioluminescence could be used as a suitable substitute for tritiated thymidine uptake as a measure of cell proliferation. The cell lines MOLT-4, HL-60, TF-1 and NFS-60 showed a strong correlation between thymidine uptake and ATP bioluminescence (p > 0.00001 for all cell types). Additionally the ATP method could detect the cytokine dependent proliferation on TF-1 and NFS-60 cells by granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) respectively. The tumour necrosis factor alpha (TNF)-induced cytotoxic effect on L-929 cells could also be accurately detected using this method. It would therefore appear to be possible to use ATP bioluminescence in the detection of cytokine activity in a number of different bioassays.

MeSH terms

  • Adenosine Triphosphate / analysis*
  • Cell Count
  • Cell Division / drug effects
  • Cell Line
  • Cells, Cultured
  • Cytotoxicity, Immunologic / drug effects
  • Cytotoxicity, Immunologic / immunology*
  • DNA Replication / drug effects
  • Firefly Luciferin
  • Granulocyte Colony-Stimulating Factor / pharmacology
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Humans
  • Leukocytes, Mononuclear / immunology*
  • Luciferases
  • Luminescent Measurements*
  • Tumor Cells, Cultured
  • Tumor Necrosis Factor-alpha / pharmacology


  • Tumor Necrosis Factor-alpha
  • Granulocyte Colony-Stimulating Factor
  • Firefly Luciferin
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Adenosine Triphosphate
  • Luciferases