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温度校正的跨上皮电阻(TEER)测量,用于量化细胞旁通透性的快速变化。

Temperature corrected transepithelial electrical resistance (TEER) measurement to quantify rapid changes in paracellular permeability.

作者信息

Blume L-F, Denker M, Gieseler F, Kunze T

机构信息

Institute of Pharmacy, University of Kiel, Germany.

出版信息

Pharmazie. 2010 Jan;65(1):19-24.

Abstract

Determining the transepithelial electrical resistance (TEER) is a widely used method to functionally analyze tight junction dynamics in cell culture models of physiological barriers. Changes in temperature are known to have strong effects on TEER and can pose problems during the process of TEER measurements in cell culture vessels, complicating comparisons of TEER data across different experiments and studies. Here, we set out to devise a strategy to obtain temperature-independent TEER values based on the physical correlation between parameters such as TEER, temperature, medium viscosity and pore size of the cell culture inserts. By measuring the impact of temperature and different electrode types on TEER measurements on Caco-2 and HPDE (normal human pancreatic ductal epithelium) monolayers, we were able to derive a mathematical method that is suitable for the correction of TEER values for temperature changes. Applying this method to raw TEER values yields temperature-corrected TEER (tcTEER) values. Validity of tcTEER was demonstrated by showing a direct correlation with permeability of monolayers as determined by flux of RITC dextran. Taken together, the mathematical solution presented here allows for a simple and accurate determination of paracellular permeability independent of temperature variation during the process of TEER recording.

摘要

测定跨上皮电阻(TEER)是一种广泛用于在生理屏障细胞培养模型中对紧密连接动力学进行功能分析的方法。已知温度变化对TEER有强烈影响,并且在细胞培养容器中进行TEER测量的过程中可能会引发问题,这使得跨不同实验和研究的TEER数据比较变得复杂。在此,我们着手设计一种策略,基于TEER、温度、培养基粘度和细胞培养插入物孔径等参数之间的物理相关性来获得与温度无关的TEER值。通过测量温度和不同电极类型对Caco-2和HPDE(正常人胰腺导管上皮)单层细胞TEER测量的影响,我们能够推导出一种适用于校正温度变化导致的TEER值的数学方法。将此方法应用于原始TEER值可得到温度校正后的TEER(tcTEER)值。通过显示tcTEER与由RITC葡聚糖通量测定的单层细胞通透性直接相关,证明了tcTEER的有效性。综上所述,本文提出的数学解决方案能够在TEER记录过程中简单而准确地测定与温度变化无关的细胞旁通透性。

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