Hydroxyethyl disulfide as an efficient metabolic assay for cell viability in vitro.
Hydroxyethyl disulfide as an efficient metabolic assay for cell viability in vitro.
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DOI：
10.1016/j.tiv.2012.01.007
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发表时间：
2012-06
影响因子：
3.2
通讯作者：
Ayene, Iraimoudi S.
中科院分区：
文献类型：
--
作者：
Li, Jie;Zhang, Donglan;Ward, Kathleen M.;Prendergast, George C.;Ayene, Iraimoudi S.
细胞活力测定有各种众所周知的实际和技术限制。现有的方法都存在非线性、背景高、协议繁琐等缺点。几种常用的四氮唑类化学物质依靠线粒体通过phenazine methosulfate （PMS）还原这些化合物形成的彩色甲酸产物。然而，灵敏度本身是有限的，因为它们的还原依赖于线粒体生物还原和PMS的细胞运输，以及四唑类化学物质的可及性。在这项研究中，我们确定了羟基乙基二硫化物（HEDS）作为一种廉价的探针，可以测量细胞代谢活性，而不需要经前质谱。在组织培养基中，HEDS以线性方式精确定量代谢活性活细胞，优于基于四氮唑和其他检测。化疗药物（顺铂、依托泊苷）、氧化剂（过氧化氢、对乙酰氨基酚）、毒素（氧化苯基胂、亚砷酸盐）或电离辐射产生的细胞毒性可通过HEDS试验快速测定。我们发现，HEDS在溶解度、膜通透性和细胞内转化为代谢报告蛋白的能力方面优于其他常用的细胞活力测定方法。我们的发现建立了一种简单、快速和低成本的方法来准确地定量活细胞。
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.
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影响因子：
3.4
作者：
Biaglow, JE;Ayene, IS;Mieyal, JJ
通讯作者：
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影响因子：
8.8
作者：
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作者：
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影响因子：
3.2
作者：
Geys, Jorina;Nemery, Benoit;Hoet, Peter H. M.
通讯作者：
Hoet, Peter H. M.