Hydroxyethyl disulfide as an efficient metabolic assay for cell viability in vitro

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• 作者：Jie Li^a ; Donglan Zhang^a ; Kathleen M. Ward^a ; George C. Prendergast^a ; ^b ; ^c ; Iraimoudi S. Ayene^a ; ^c ; ^{ayenei@mlhs.org}
• 关键词：G6PD ; glucose-6-phosphate dehydrogenase ; HEDS ; hydroxyethyldisulfide ; OPPC ; oxidative pentose phosphate cycle ; ME ; mercaptoethanol ; DTNB ; dithiobisnitrobenzoicacid ; HTS ; high throughput screening
• 刊名：Toxicology in Vitro
• 出版年：2012
• 出版时间：June, 2012
• 年：2012
• 卷：26
• 期：4
• 页码：603-612
• 全文大小：524 K

文摘

Cell viability assays have a variety of well known practical and technical limitations. All the available approaches have disadvantages, such as non-linearity, high background and cumbersome protocols. Several commonly used tetrazolium chemicals rely upon generation of a colored formazan product formed by mitochondrial reduction of these compounds via phenazine methosulfate (PMS). However, sensitivity is inherently limited because their reduction relies on mitochondrial bioreduction and cellular transport of PMS, as well as accessibility to tetrazolium chemicals. In this study, we identify hydroxethyldisulfide (HEDS) as an inexpensive probe that can measure cellular metabolic activity without the need of PMS. In tissue culture medium, HEDS accurately quantitated metabolically active live cells in a linear manner superior to tetrazolium based and other assays. Cell toxicity produced by chemotherapeutics (cisplatin, etoposide), oxidants (hydrogen peroxide, acetaminophen), toxins (phenyl arsine oxide, arsenite) or ionizing radiation was rapidly determined by the HEDS assay. We found that HEDS was superior to other commonly used assays for cell viability determinations in its solubility, membrane permeability, and intracellular conversion to a metabolic reporter that is readily transported into the extracellular medium. Our findings establish the use of HEDS in a simple, rapid and low cost assay to accurately quantify viable cells.