Department of Electronics and Telecommunications (DET), Politecnico di Torino, Turin, Italy.
Institute of Biostructure and Bioimaging (CNR), Molecular Biotechnology Center (MBC), Turin, Italy.
Talanta. 2021 Jan 15;222:121441. doi: 10.1016/j.talanta.2020.121441. Epub 2020 Aug 25.
Interest in impedance-based cellular assays is rising due to their remarkable advantages, including label-free, low cost, non-invasive, non-destructive, quantitative and real-time monitoring. In order to test their potential in cancer treatment decision and early detection of chemoresistance, we devised a new custom-made impedance measuring system based on electric cell-substrate impedance sensing (ECIS), optimized for long term impedance measurements. This device was employed in a proof of concept cell culture impedance analysis for the characterization of chemo-resistant colon cancer cells. Doxorubicin-resistant HT-29 cells were used for this purpose and monitored for 140 h. Analysis of impedance-based curves reveal different trends from chemo-sensitive and chemo-resistant cells. An impedance-based cytoxicity assay with different concentrations of doxorubicin was also performed using ECIS. The obtained results confirm the feasibility of ECIS in the study of drug resistance and show promises for studies of time-dependent factors related to physiological and behavioral changes in cells during resistance acquisition. The methodology presented herein, allows the continuous monitoring of cells under normal culture conditions as well as upon drug exposure. The ECIS device used, sets the basis for high-throughput early detection of resistance to drugs, administered in the clinical practice to cancer patients, and for the screening of new drugs in vitro, on patient-derived cells.
由于其显著的优势，基于阻抗的细胞检测法引起了人们的兴趣，包括无标记、低成本、非侵入性、非破坏性、定量和实时监测。为了测试它们在癌症治疗决策和早期耐药性检测中的潜力，我们设计了一种新的基于电细胞-基质阻抗传感（ECIS）的定制阻抗测量系统，该系统经过优化，可进行长期阻抗测量。该设备用于概念验证细胞培养阻抗分析，以表征化疗耐药的结肠癌细胞。为此，使用多柔比星耐药的 HT-29 细胞，并监测 140 小时。基于阻抗的曲线分析揭示了化疗敏感和耐药细胞的不同趋势。还使用 ECIS 进行了不同浓度多柔比星的基于阻抗的细胞毒性测定。获得的结果证实了 ECIS 在耐药性研究中的可行性，并为研究与细胞获得耐药性过程中生理和行为变化相关的时间依赖性因素提供了前景。本文提出的方法允许在正常培养条件下以及药物暴露下连续监测细胞。所使用的 ECIS 设备为临床癌症患者用药的耐药性的高通量早期检测以及基于患者来源细胞的新药物的体外筛选奠定了基础。
