High-Throughput Viability Assay Using an Autonomously Bioluminescent Cell Line with a Bacterial Lux Reporter
High-Throughput Viability Assay Using an Autonomously Bioluminescent Cell Line with a Bacterial Lux Reporter
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DOI：
10.1177/2211068214560608
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发表时间：
2015-04-01
期刊：
影响因子：
--
通讯作者：
Zheng, Wei
中科院分区：
文献类型：
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作者：
Class, Bradley;Thorne, Natasha;Zheng, Wei
细胞活力测定广泛用于确定细胞健康、评价生长条件和评估化合物细胞毒性。大多数现有的测定是终点测定，其中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机制。理想的细胞活力测定允许在动力学上测定化合物毒性，而不必过早终止测定。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HEK 293细胞系优化并验证了无试剂添加的细胞活力测定。该细胞活力测定可用于实时、长期测量活细胞中的化合物细胞毒性，与化合物孵育24、48- 72或96小时后，信号与基础比率为20至200倍，Z因子为0.6。我们还发现，九种细胞毒性化合物的效力与其他四种常用的细胞活力测定法测得的效力相关。结果表明，使用HEK 293（lux）自发光细胞系的动态细胞活力试验可用于化合物细胞毒性的高通量评价。
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.