Abstract
The commonly used trypan blue dye exclusion method and other modified cell viability methods, such as fluorescein dye and tetrazolium dye exclusion, artificially introduce toxic chemicals to cells and, thus, alter cellular organelles when measuring cell viability. Therefore, cell viability could be affected by the processes currently used to observe viability. In this study, the cell viability of Chinese hamster ovary (CHO) cells was measured by simply counting attached cells after the cultured CHO cells were attached on a Concanavalin A (Con A) substrate. The efficiency of cell attachment to Con A surfaces was different for live and dead cells allowing the cell viability of CHO cells to be measured without any chemical modifications to the cells.
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Jones, K. H. and J. A. Senft (1985) An improved method to determine cell viability by simultaneous staining with fluorescein diacetate-propidium iodide. J. Histochem. Cytochem. 33: 77–79.
Kwok, A. K., C. K. Yeung, T. Y. Lai, K. P. Chan, and C. P. Pang (2004) Effects of trypan blue on cell viability and gene expression in human retinal pigment epitherial cells. Brit. J. Ophthalmol. 88: 1590–1594.
Foglieni, C., C. Meoni, and A. M. Davalli (2001) Fluorescent dyes for cell viability: An application on prefixed conditions. Histochem. Cell Biol. 115: 223–229.
Legrand, C., J. M. Bour, C. Jacob, J. Capiaumont, A. Martial, A. Marc, M. Wudtke, G. Kretzmer, C. Demanqel, and D. Duval (1992) Lactate dehydrogenase (LDH) activity of the number of the cultured eukaryotic cells as marker of the dead cells in the medium. J. Biotechnol. 25: 231–243.
Berridge, M. V., P. M. Herst, and A. S. Tan (2005) Tetrazolium dyes as tools in cell biology: New insights into their cellular reduction. Biotechnol. Annu. Rev. 11: 127–152.
Ishiyama, M., H. Tominaga, M. Shiga, K. Sasamoto, Y. Ohkura, and K. Ueno (1996) A combined assay of cell viability and in vitro cytotoxicity with a highly water-soluble tetrazolium salt, neutral red and crystal violet. Biol. Pharm. Bull. 19: 1518–1520.
O’Brien, J., I Wilson, T. Orton, and F. Pognan (2000) Investigation of the alalmar blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur. J. Biochem. 267: 5421–5426.
Ahmed, S. A., R. M. Gogal Jr, and J. E. Walsh (1994) A new rapid and simple non-radioactive assay to monitor and determine the proliferation of lymphocytes: An alternative to [3H] thymidine incorporation assay. J. Immunol. Methods 170: 211–224.
Alberts, B., A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter (2002) Molecular Biology of the Cell. 4th ed., Garland Science, NY, USA.
Zanetta, J. -P., S. Kuchler, S. Lehmann, A. Badache, S. Maschke, D. Thomas, P. Dufourcq, and G. Vincendon (1992) Glycoproteins and lectins in cell adhesion and cell recognition processes. Histochem. J. 24: 791–804.
Caro-Maldonado, A. and C. M. Pinedo (2011) Dying for something to eat: How cells respond to starvation. Open Cell Signal. J. 3: 42–51.
Hardie, D. G. (2007) AMP-activated/SNF1 protein kinase: Conserved guardians of cellular energy Nat. Rev. Mol. Cell Bio. 8: 774–785.
Wang, X. and C. G. Proud (2009) Nutrient control of TORC1, a cell-cycle regulator. Trends Cell Biol. 19: 260–267.
Vetri, V., G. Ossato, V. Militello, M. A. Digman, M. Leone, and E. Gratton (2011) Fluctuation methods to study protein aggregation in live cells: Concanavalin A oligomers formation. Biophys. J. 100: 774–783.
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Ahn, J., Park, J., Kim, YG. et al. Label-free measurement of cell viability via counting cells attached on affinity substrates. Biotechnol Bioproc E 19, 257–261 (2014). https://doi.org/10.1007/s12257-013-0556-1
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DOI: https://doi.org/10.1007/s12257-013-0556-1