Adan Aysun, Kiraz Yağmur, Baran Yusuf
İzmir Institute of Technology, Department of Molecular Biology and Genetics, İzmir, Turkey.
Curr Pharm Biotechnol. 2016;17(14):1213-1221. doi: 10.2174/1389201017666160808160513.
Cell viability is defined as the number of healthy cells in a sample and proliferation of cells is a vital indicator for understanding the mechanisms in action of certain genes, proteins and pathways involved cell survival or death after exposing to toxic agents. Generally, methods used to determine viability are also common for the detection of cell proliferation. Cell cytotoxicity and proliferation assays are generally used for drug screening to detect whether the test molecules have effects on cell proliferation or display direct cytotoxic effects. Regardless of the type of cell-based assay being used, it is important to know how many viable cells are remaining at the end of the experiment. There are a variety of assay methods based on various cell functions such as enzyme activity, cell membrane permeability, cell adherence, ATP production, co-enzyme production, and nucleotide uptake activity. These methods could be basically classified into different categories: (I) dye exclusion methods such as trypan blue dye exclusion assay, (II) methods based on metabolic activity, (III) ATP assay, (IV) sulforhodamine B assay, (V) protease viability marker assay, (VI) clonogenic cell survival assay, (VII) DNA synthesis cell proliferation assays and (V) raman micro-spectroscopy. In order to choose the optimal viability assay, the cell type, applied culture conditions, and the specific questions being asked should be considered in detail. This particular review aims to provide an overview of common cell proliferation and cytotoxicity assays together with their own advantages and disadvantages, their methodologies, comparisons and intended purposes.
细胞活力定义为样本中健康细胞的数量，细胞增殖是了解某些基因、蛋白质和参与细胞在接触毒性剂后存活或死亡的信号通路作用机制的重要指标。一般来说，用于确定活力的方法也常用于检测细胞增殖。细胞毒性和增殖测定通常用于药物筛选，以检测测试分子是否对细胞增殖有影响或显示直接细胞毒性作用。无论使用何种基于细胞的测定类型，了解实验结束时剩余的活细胞数量都很重要。基于各种细胞功能，如酶活性、细胞膜通透性、细胞黏附、ATP产生、辅酶产生和核苷酸摄取活性，有多种测定方法。这些方法基本上可分为不同类别：（I）染料排斥法，如台盼蓝染料排斥试验；（II）基于代谢活性的方法；（III）ATP测定；（IV）磺基罗丹明B测定；（V）蛋白酶活力标记测定；（VI）克隆形成细胞存活测定；（VII）DNA合成细胞增殖测定和（V）拉曼显微光谱法。为了选择最佳的活力测定方法，应详细考虑细胞类型、应用的培养条件以及所提出的具体问题。本综述旨在概述常见的细胞增殖和细胞毒性测定方法，以及它们各自的优缺点、方法、比较和预期用途。
