Principal investigator: Iraimoudi S. Ayene, PhD
Cell growth assays used in the laboratory have many widely known, practical and technical limitations, including non-linearity, high background and cumbersome protocols. In particular, commonly used assays using tetrazolium salts to generate a colored product lack sensitivity and accuracy due to reliance on mitochondrial bioreduction and other factors.
Addressing these limitations, LIMR researchers have developed a fast, accurate and inexpensive assay suitable for measuring cell growth and viability in tissue culture settings. This assay, termed CellCountEZ, uses a nontoxic detector compound that does not compromise cell viability itself, enabling experimental and bioreactor uses in which periodic longitudinal measurements are desired.
CellCountEZ quickly and accurately quantitates relative cell number in a highly linear manner, based on colorimetric detection of beta-mercaptoethanol produced by metabolic reduction of the dithiol reporter compound hydroxyethyl disulfide. CellCountEZ also can be used to quickly quantify the relative number of viable cells remaining in tissue culture after toxic treatments (e.g. chemotherapeutics, oxidants, radiation).
We have found this test to be superior to other tissue culture assays in its ability to rapidly and accurately determine relative cell number and viability in a nontoxic fashion.
Biotechnology companies and biology laboratories have a universal need for accurate measurements of relative cell growth and viability in tissue culture media. CellCountEZ offers an assay system that is rapid, accurate, highly linear and inexpensive. It is amenable to any eukaryotic cell system. The detection compound used in this patented system is non-toxic, enabling its use in bioreactors to measure cell viability and growth longitudinally to the highest cell densities without loss of linear response.
These features of CellCountEZ render it useful in tissue culture settings used in biology laboratories and biotechnology companies.
Intellectual property position
Method of determining cell number or viability using hydroxyethyldisulfide: U.S. patent No. 8,697,391 (issued 15 April, 2014).
Li J, Zhang D, Ward KM, Prendergast GC, Ayene IS. (2012). Hydroxyethyl disulfide as an efficient metabolic assay for cell viability in vitro. Toxicol In Vitro, 26;603-12.