Division of Solid Organ Transplantation, Department of Surgery, University of Rochester Medical Center, Rochester, NY 14623, USA.
Anal Biochem. 2012 Oct 1;429(1):32-8. doi: 10.1016/j.ab.2012.07.006. Epub 2012 Jul 14.
We report the use of resazurin (AlamarBlue) dye in a robust assay for cell viability of primary cells. Human mononuclear cells were used here for immunological studies, but the method can be applied to monitor reduction potential of any living cell. Reduction of AlamarBlue dye is widely used in several commercial assays of cell viability. Although it is fast and easy with immortal cell lines, the method is impractical for the primary cells due to their slower metabolic activity. We propose that the viability of human primary cells can be determined with AlamarBlue by monitoring the increase in fluorescence intensity in a matter of a few hours. In the presence of AlamarBlue, the dynamic increase in cellular reduction capacity is linear for several hours or, alternatively, the assay can be repeated to monitor the viability at any time point of cell culture. In addition to testing cellular growth rates and cytotoxicity, the application can be used to compare sample quality of cells that have been frozen or represent a pool of multiple donors. This application of the AlamarBlue cell viability assay is simple, rapid, and cost-effective, and therefore it is also well suited for high-throughput studies.
我们报告了使用 Resazurin（AlamarBlue）染料在用于原代细胞活力的可靠测定中的应用。这里使用人单核细胞进行免疫学研究，但是该方法可用于监测任何活细胞的还原电势。AlamarBlue 染料的还原广泛用于几种细胞活力的商业测定中。尽管对于永生化细胞系来说它既快速又简单，但由于原代细胞的代谢活性较慢，因此该方法并不适用于原代细胞。我们提出，通过在数小时内监测荧光强度的增加，可以用 AlamarBlue 测定人原代细胞的活力。在 AlamarBlue 存在下，细胞还原能力的动态增加在数小时内呈线性，或者可以重复该测定以在细胞培养的任何时间点监测活力。除了测试细胞生长速度和细胞毒性外，该应用还可用于比较已冷冻或代表多个供体池的细胞的样品质量。AlamarBlue 细胞活力测定的这种应用简单、快速且具有成本效益，因此也非常适合高通量研究。
