Opp Daniel, Wafula Brian, Lim Jennifer, Huang Eric, Lo Jun-Chih, Lo Chun-Min
Department of Physics, University of South Florida, Tampa, FL 33620-5700, United States.
Biosens Bioelectron. 2009 Apr 15;24(8):2625-9. doi: 10.1016/j.bios.2009.01.015. Epub 2009 Jan 23.
In vitro assessment of cytotoxicity based on electrochemical impedance spectroscopy (EIS) needs more quantitative methods to analyze the alteration of cell morphology and motility, and hence the potential risk to human health. Here, we applied electric cell-substrate impedance sensing (ECIS) to evaluate dose-dependent responses of human umbilical vein endothelial cells exposed to cytochalasin B. To detect subtle changes in cell morphology, the frequency-dependent impedance data of the cell monolayer were measured and analyzed with a theoretical cell-electrode model. To detect the alternation of cell micromotion in response to cytochalasin B challenge, time-series impedance fluctuations of cell-covered electrodes were monitored and the values of power spectrum, variance, and variance of the increments were calculated to verify the difference. While a dose-dependent relationship was generally observed from the overall resistance of the cell monolayer, the analysis of frequency-dependent impedance and impedance fluctuations distinguished cytochalasin B levels as low as 0.1 microM. Our results show that cytochalasin B causes a decrease of junctional resistance between cells, an increase of membrane capacitance, and the reduction in micromotion.
基于电化学阻抗谱(EIS)的细胞毒性体外评估需要更多定量方法来分析细胞形态和运动性的变化,进而分析对人类健康的潜在风险。在此,我们应用细胞-基质阻抗传感(ECIS)来评估暴露于细胞松弛素B的人脐静脉内皮细胞的剂量依赖性反应。为了检测细胞形态的细微变化,我们用理论细胞-电极模型测量并分析了细胞单层的频率依赖性阻抗数据。为了检测细胞对细胞松弛素B刺激的微运动变化,我们监测了细胞覆盖电极的时间序列阻抗波动,并计算了功率谱、方差和增量方差的值以验证差异。虽然通常从细胞单层的总电阻观察到剂量依赖性关系,但频率依赖性阻抗和阻抗波动分析能够区分低至0.1微摩尔的细胞松弛素B水平。我们的结果表明,细胞松弛素B会导致细胞间连接电阻降低、膜电容增加以及微运动减少。