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使用 ECIS TEER96 在 96 孔 Transwell 中自动测量跨上皮电阻 (TEER):单次和多次时间点评估。

Automated measurement of transepithelial electrical resistance (TEER) in 96-well transwells using ECIS TEER96: Single and multiple time point assessments.

机构信息

Division of Preclinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, 9800 Medical Center Dr, Rockville, MD 20850.

Applied BioPhysics, 185 Jordan Rd, Troy, NY 12180.

出版信息

SLAS Technol. 2024 Feb;29(1):100116. doi: 10.1016/j.slast.2023.10.008. Epub 2023 Nov 2.

Abstract

Transepithelial electrical resistance (TEER) is a widely used technique for quantifying the permeability of epithelial and endothelial cell layers. However, traditional methods of measuring TEER are limited to single timepoint measurements and can subject cells to an altered environment during the measurement. Here, we assessed the validity of TEER measurements by the ECIS TEER96 device, which is designed to take continuous TEER measurements of a cell culture system in a standard laboratory incubator. We found that the instrument accurately measures TEER across TEER values ranging from 10 to 2050 Ω*cm and is more accurate than the manual epithelial voltohmmeter electrode at high TEER values. Furthermore, the high-resolution measurements provided by the device allowed for a unique insight into the mechanisms and kinetics of cells in vitro. To demonstrate the continuous measurement capability of the device, we tracked the formation of an MDCKI cell monolayer until TEER plateaued. Furthermore, we treated Caco-2 monolayers with different concentrations of DMSO and the antimicrobial and surfactant compound benzethonium chloride to measure disruptions to barrier integrity. Treatment of both compounds resulted in concentration-dependent loss of barrier integrity. Our results suggest that the ECIS TEER96 device is a reliable and convenient option for measuring TEER in cell cultures and can provide valuable insights into the behavior of cells in vitro. This technology will be especially useful for increasing throughput of drug permeability assays, inflammation studies, and gaining better understanding of disease states in a cell culture system.

摘要

跨上皮电阻(TEER)是一种广泛用于量化上皮和内皮细胞层通透性的技术。然而,传统的 TEER 测量方法仅限于单点测量,并且在测量过程中会使细胞处于改变的环境中。在这里,我们评估了 ECIS TEER96 设备测量 TEER 的有效性,该设备旨在在标准实验室培养箱中对细胞培养系统进行连续的 TEER 测量。我们发现该仪器可以准确测量 TEER 值在 10 到 2050 Ω*cm 之间的 TEER 值,并且在高 TEER 值下比手动上皮电压计电极更准确。此外,该设备提供的高分辨率测量使我们能够深入了解细胞在体外的机制和动力学。为了展示设备的连续测量能力,我们跟踪了 MDCKI 细胞单层的形成,直到 TEER 达到稳定。此外,我们用不同浓度的 DMSO 和抗菌和表面活性剂化合物苯扎氯铵处理 Caco-2 单层,以测量对屏障完整性的破坏。两种化合物的处理都导致了屏障完整性的浓度依赖性丧失。我们的结果表明,ECIS TEER96 设备是测量细胞培养物中 TEER 的可靠且方便的选择,并且可以提供有关细胞在体外行为的有价值的见解。这项技术对于增加药物通透性测定、炎症研究的通量以及更好地理解细胞培养系统中的疾病状态将特别有用。

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