Class Bradley, Thorne Natasha, Aguisanda Francis, Southall Noel, McKew John C, Zheng Wei
National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA
J Lab Autom. 2015 Apr;20(2):164-74. doi: 10.1177/2211068214560608. Epub 2014 Dec 1.
Cell viability assays are extensively used to determine cell health, evaluate growth conditions, and assess compound cytotoxicity. Most existing assays are endpoint assays, in which data are collected at one time point after termination of the experiment. The time point at which toxicity of a compound is evident, however, depends on the mechanism of that compound. An ideal cell viability assay allows the determination of compound toxicity kinetically without having to terminate the assay prematurely. We optimized and validated a reagent-addition-free cell viability assay using an autoluminescent HEK293 cell line that stably expresses bacterial luciferase and all substrates necessary for bioluminescence. This cell viability assay can be used for real-time, long-term measurement of compound cytotoxicity in live cells with a signal-to-basal ratio of 20- to 200-fold and Z-factors of ~0.6 after 24-, 48- 72-, or 96-h incubation with compound. We also found that the potencies of nine cytotoxic compounds correlated well with those measured by four other commonly used cell viability assays. The results demonstrated that this kinetic cell viability assay using the HEK293(lux) autoluminescent cell line is useful for high-throughput evaluation of compound cytotoxicity.
细胞活力测定被广泛用于确定细胞健康状况、评估生长条件以及评估化合物的细胞毒性。大多数现有的测定是终点测定，即在实验终止后的一个时间点收集数据。然而，化合物毒性明显的时间点取决于该化合物的作用机制。理想的细胞活力测定能够动态地确定化合物毒性，而无需过早终止测定。我们使用稳定表达细菌荧光素酶和生物发光所需的所有底物的自发光HEK293细胞系，优化并验证了一种无需添加试剂的细胞活力测定方法。这种细胞活力测定可用于实时、长期测量活细胞中化合物的细胞毒性，在与化合物孵育24、48、72或96小时后，信号与本底比为20至200倍，Z因子约为0.6。我们还发现，九种细胞毒性化合物的效力与通过其他四种常用细胞活力测定方法测得的效力具有良好的相关性。结果表明，这种使用HEK293(lux)自发光细胞系的动态细胞活力测定对于化合物细胞毒性的高通量评估是有用的。
