Validation of the cytosensor for in vitro cytotoxicity studies
Validation of the cytosensor for in vitro cytotoxicity studies
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DOI：
10.1016/s0887-2333(97)00009-x
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发表时间：
1997-06-01
影响因子：
3.2
通讯作者：
Eldefrawi, AT
中科院分区：
文献类型：
--
作者：
Cao, CJ;Mioduszewski, RJ;Eldefrawi, AT
使用细胞传感器（TM）微生理仪连续监测暴露于10种药物时人肝细胞系ATCC-CCL-13代谢率的扰动。比较暴露于同一浓度24小时或连续增加浓度4小时的影响，以及药物去除后的恢复情况。用对乙酰氨基酚（扑热息痛）和乙醇建立测定方案，确定药物作用的可逆性。暴露24小时后，所有药物都产生了浓度和时间依赖性的酸化速率降低，这可能是由于活细胞数量减少和/或活细胞代谢率降低。以不可逆抑制酸化速率的程度作为细胞死亡的指标，10种药物的IC50值与仅在活细胞中荧光的钙黄素AM荧光法（r = 0.991）和[H-3]胸苷摄取法（r = 0.976）在同一细胞系中产生的IC50值相当。从细胞传感器获得的24小时暴露的IC50值与10种药物公布的人类致死血液浓度之间也有很好的相关性（r = 0.958）。与其他体外测定法相比，这种新方法的一个优点是，它允许确定可逆变化变为不可逆的时间点。1997爱思唯尔科学有限公司
The Cytosensor(TM) microphysiometer was used to continuously monitor perturbations in metabolic rates of the human liver cell line ATCC-CCL-13 when exposed to each of 10 drugs. The effects of exposure to one concentration for 24 hr or to sequential increasing concentrations for 4 hr, and recovery after drug removal, were compared. Paracetamol (acetaminophen) and ethanol were used to establish the assay protocols and determine reversibility of drug effect. All drugs produced concentration- and time-dependent reduction in acidification rate following 24 hr exposure, which may be due to decreased number of viable cells and/or lowered metabolic rates of the live cells. The degree of irreversible inhibition of acidification rate was used as an index of cell death and the IC50 values for the 10 drugs were comparable to those produced in the same cell line by a fluorescence assay using Calcein AM stain (r = 0.991), that fluoresces only in live cells, as well as the [H-3]thymidine uptake assay (r = 0.976). There was also excellent correlation (r = 0.958) between IC50 values of 24 hr exposure obtained from the Cytosensor with the 10 drugs and their published human lethal blood concentrations. An advantage of this new methodology over other in vitro assays is that it allows the determination of time points at which reversible change becomes irreversible. (C) 1997 Elsevier Science Ltd.