Neutral red uptake assay for the estimation of cell viability/cytotoxicity
Neutral red uptake assay for the estimation of cell viability/cytotoxicity
复制标题
DOI：
10.1038/nprot.2008.75
复制
发表时间：
2008-01-01
期刊：
影响因子：
14.8
通讯作者：
Zurita, Jorge L.
中科院分区：
文献类型：
--
作者：
Repetto, Guillermo;del Peso, Ana;Zurita, Jorge L.
中性红摄取试验提供了对培养物中活细胞数量的定量估计。它是最常用的细胞毒性测试之一，具有许多生物医学和环境应用。它是基于活细胞结合和结合溶酶体中的超生命染料中性红的能力。大多数原代细胞和来自不同来源的细胞系都可以成功地使用。细胞接种在96孔组织培养板中，并在适当的时间内进行处理。然后在含有中性红的培养液中培养2小时。随后对细胞进行清洗，在每个孔中提取染料，并使用分光光度计读取吸光度。该程序比其他细胞毒性试验(四氮唑盐、酶渗漏或蛋白质含量)更便宜、更敏感。一旦细胞经过处理，检测可以在3小时内完成。
The neutral red uptake assay provides a quantitative estimation of the number of viable cells in a culture. It is one of the most used cytotoxicity tests with many biomedical and environmental applications. It is based on the ability of viable cells to incorporate and bind the supravital dye neutral red in the lysosomes. Most primary cells and cell lines from diverse origin may be successfully used. Cells are seeded in 96-well tissue culture plates and are treated for the appropriate period. The plates are then incubated for 2 h with a medium containing neutral red. The cells are subsequently washed, the dye is extracted in each well and the absorbance is read using a spectrophotometer. The procedure is cheaper and more sensitive than other cytotoxicity tests (tetrazolium salts, enzyme leakage or protein content). Once the cells have been treated, the assay can be completed in < 3 h.