Discriminative cytotoxicity assessment based on various cellular damages
Discriminative cytotoxicity assessment based on various cellular damages
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DOI：
10.1016/j.toxlet.2008.10.006
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发表时间：
2009-01-10
影响因子：
3.5
通讯作者：
Jeong, Damon
中科院分区：
文献类型：
--
作者：
Kim, Hyunsoo;Yoon, Sung Chul;Jeong, Damon
目前有几种测定法可用于评估细胞毒性，包括台盼蓝排除法、乳酸脱氢酶（LDH）释放法和3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四唑（MTT）还原测定法。台盼蓝排斥和LDH释放试验适用于评价细胞膜损伤，MTT比色试验可用于测量细胞相关的还原能力。由于这些试验是随机使用的，以评估细胞损伤的程度，我们在此建议，该试验应根据主要的细胞情况进行选择。这可以通过使用具有不同还原状态的多种细胞类型、外源性和内源性氧化应激源以及几种不同的氧化/还原分子来确定。尽管台盼蓝排除法和释放LDH测定法已被证明对于评估具有膜损伤的坏死和凋亡细胞死亡有用，但LDH测定法不适合测量没有膜损伤的各种细胞的数量。此外，当细胞用外源性和内源性氧化应激物处理时，MTT还原显示出对在窄范围内转移到更氧化的细胞环境而不损失膜完整性敏感，并且这种效果以线性方式增加，依赖于含有各种生理还原剂、小还原分子（NADPH和GSH）和人工DTT还原剂。最后，我们注意到MTT试验可用于测定细胞还原状态的小规模振荡和线粒体功能活性的变化，但不用于评价具有较高细胞还原能力的细胞的细胞毒性。总之，本研究的结果表明，应通过考虑各种刺激的细胞条件与测定系统的基本原理之间的相关性，选择不同的细胞毒性试验工具。(C)2008爱思唯尔爱尔兰有限公司保留所有权利。
There are several assays currently available for the assessment of cell cytotoxicity, including trypan blue exclusion, lactate dehydrogenase (LDH) release, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assays. Trypan blue exclusion and LDH release assays are appropriate for evaluating cell membrane damage and a colorimetric MTT assay is available for measuring mitochondrial-related reduction capacity. As these assays were randomly utilized to assess the extent of cell damage, we suggest herein that the assay should be selected in accordance with the prevailing cellular situation. This can be determined by using a variety of cell types with differing reduction status, exogenous and endogenous oxidative stressors, and several different oxidized/reduced molecules. Although the trypan blue exclusion and released LDH assay have proven useful for assessments of necrotic and apoptotic cell death with membrane damage, the LDH assay is not appropriate for the measurement of the number of varied cells without membrane damage. In addition, when the cells were treated with exogenous and endogenous oxidative stressors, MTT reduction was shown to be sensitive to a shift to a more oxidizing cellular environment within a narrow range without loss of membrane integrity, and this effect increased in a linear fashion, dependent on the dosage of cytosolic extracts containing various physiological reductants, small reductive molecules (NADPH and GSH), and artificial DTT reducing agent. Finally, we noted that the MTT assay is available for the determination of small-scale oscillations in cellular reduction status and changes in mitochondrial functional activity, but not for evaluating the cytotoxicity of cells with a higher cellular reduction capacity. Altogether, the findings of this study indicate that tools for the testing of cytotoxicity should be selected differently by considering the correlation between the cellular conditions for various stimuli and the principle underlying the assay system. (C) 2008 Elsevier Ireland Ltd. All rights reserved.