使用ECIS TEER96自动测量96孔Transwell中的跨上皮电阻(TEER):单时间点和多时间点评估。

IF 2.5 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS SLAS Technology Pub Date : 2024-02-01 DOI:10.1016/j.slast.2023.10.008
Jacob Schimetz , Pranav Shah , Charles Keese , Chris Dehnert , Michael Detweiler , Sam Michael , Catherine Toniatti-Yanulavich , Xin Xu , Elias C. Padilha
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引用次数: 0

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,测量TEER的传统方法仅限于单个时间点测量,并且在测量过程中可能使细胞受到改变的环境。在这里,我们通过ECIS TEER96设备评估了TEER测量的有效性,该设备旨在对标准实验室培养箱中的细胞培养系统进行连续的TEER测量。我们发现,该仪器在10-2050Ω*cm[2]的TEER值范围内准确测量TEER,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结果使我们能够独特地了解体外细胞的机制和动力学。为了证明该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稳定。此外,我们用不同浓度的二甲基亚砜和抗菌和表面活性剂化合物苯扎氯铵处理Caco-2单层,以测量屏障完整性的破坏。两种化合物的处理导致屏障完整性的浓度依赖性损失。我们的研究结果表明,ECIS TEER96设备是一种可靠而方便的测量细胞培养物中TEER的选择,可以为体外细胞的行为提供有价值的见解。这项技术将特别有助于提高药物渗透性测定、炎症研究的吞吐量,并更好地了解细胞培养系统中的疾病状态。
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Automated measurement of transepithelial electrical resistance (TEER) in 96-well transwells using ECIS TEER96: Single and multiple time point assessments

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*cm2 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.

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来源期刊
SLAS Technology
SLAS Technology Computer Science-Computer Science Applications
CiteScore
6.30
自引率
7.40%
发文量
47
审稿时长
106 days
期刊介绍: SLAS Technology emphasizes scientific and technical advances that enable and improve life sciences research and development; drug-delivery; diagnostics; biomedical and molecular imaging; and personalized and precision medicine. This includes high-throughput and other laboratory automation technologies; micro/nanotechnologies; analytical, separation and quantitative techniques; synthetic chemistry and biology; informatics (data analysis, statistics, bio, genomic and chemoinformatics); and more.
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