Hajek Kathrin, Wegener Joachim
Institut für Analytische Chemie, Chemo- und Biosensorik, Universität Regensburg, Universitätsstr. 31, 93053 Regensburg, Germany.
Institut für Analytische Chemie, Chemo- und Biosensorik, Universität Regensburg, Universitätsstr. 31, 93053 Regensburg, Germany.
Exp Cell Res. 2017 Feb 1;351(1):121-126. doi: 10.1016/j.yexcr.2017.01.003. Epub 2017 Jan 10.
The transepithelial or -endothelial electrical resistance (TEER) is a very common and routinely recorded parameter describing the expression of barrier-forming cell-cell contacts (tight junctions) in quantitative terms. To determine TEER, barrier-forming cell monolayers are cultured on porous filter supports that separate two fluid compartments. The frequency-dependent impedance of the cell layer is then recorded and analyzed by means of equivalent circuit modelling providing TEER and the cell layer capacitance. The latter serves as a quantitative indicator for membrane topography. When cells are co-cultured on opposite sides of such a porous support to model more complex biological barriers, TEER readings will integrate over both cell layers and the individual contributions are not assessable. This study describes the modification of commonly used porous filter inserts by coating their backside with a thin gold-film. When this gold-film is used as an additional electrode, both cell layers can be studied separately by impedance analysis. The electrical parameters of either cell layer are assessable independently by switching between different electrode combinations. The performance of this new approach is illustrated and documented by experiments that (i) follow the de novo formation of cell junctions between initially suspended cells and (ii) the manipulation of mature cell-cell junctions by cytoskeleton-active drugs. Both assays confirm that both cell layers are monitored entirely independently.
跨上皮或跨内皮电阻（TEER）是一个非常常见且常规记录的参数，用于定量描述形成屏障的细胞间接触（紧密连接）的表达。为了测定TEER，将形成屏障的细胞单层培养在分隔两个液体隔室的多孔滤膜支架上。然后通过等效电路建模记录并分析细胞层的频率依赖性阻抗，从而得出TEER和细胞层电容。后者作为膜拓扑结构的定量指标。当细胞在这种多孔支架的相对两侧共培养以模拟更复杂的生物屏障时，TEER读数将整合两个细胞层的情况，无法评估各个细胞层的单独贡献。本研究描述了通过在常用多孔滤膜插入物的背面涂覆一层薄金膜来对其进行改进。当将该金膜用作附加电极时，可通过阻抗分析分别研究两个细胞层。通过在不同电极组合之间切换，可以独立评估任一细胞层的电参数。通过以下实验说明并记录了这种新方法的性能：（i）跟踪最初悬浮细胞之间细胞连接的从头形成，以及（ii）用细胞骨架活性药物操纵成熟的细胞间连接。这两种测定方法均证实两个细胞层是完全独立监测的。
