Discrepancies between metabolic activity and DNA content as tool to assess cell proliferation in cancer research.
Cell proliferation is a critical and frequently studied feature of molecular biology in cancer research. Therefore, various assays are available using different strategies to measure cell proliferation. Metabolic assays such as AlamarBlue, water-soluble tetrazolium salt and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, which were originally developed to determine cell toxicity, are used to assess cell numbers. Additionally, proliferative activity can be determined by quantification of DNA content using fluorophores such as CyQuant and PicoGreen. Referring to data published in high ranking cancer journals, these assays were applied in 945 publications over the past 14 years to examine the proliferative behaviour of diverse cell types. In these studies, however, mainly metabolic assays were used to quantify changes in cell growth yet these assays may not accurately reflect cellular proliferation rates due to a miscorrelation of metabolic activity and cell number. Testing this hypothesis, we compared the metabolic activity of different cell types, human cancer cells and primary cells, over a time period of 4 days using AlamarBlue and the fluorometric assays CyQuant and PicoGreen to determine their DNA content. Our results show certain discrepancies in terms of over-estimation of cell proliferation with respect to the metabolic assay in comparison to DNA binding fluorophores.
Quent
Verena M C
VM
Regenerative Medicine, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland, Australia.
Loessner
Daniela
D
Friis
Thor
T
Reichert
Johannes C
JC
Hutmacher
Dietmar W
DW
D000368:Aged; D001681:Biological Assay; D035843:Biomedical Research / Q000379:methods*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004273:DNA, Neoplasm / Q000378:metabolism*; D005260:Female; D006801:Humans; D009369:Neoplasms / Q000378:metabolism* / Q000473:pathology*; D010078:Oxazines / Q000378:metabolism; D011642:Publications; D013997:Time Factors; D014966:Xanthenes / Q000378:metabolism
D004273:DNA, Neoplasm; D010078:Oxazines; D014966:Xanthenes; C005843:resazurin