Wegener Joachim, Abrams Dimitri, Willenbrink Wolfgang, Galla Hans-Joachim, Janshoff Andreas
Institüt für Biochemie, Westfälische Wilhelms-Universität Münster, Münster, Germany.
Biotechniques. 2004 Oct;37(4):590, 592-4, 596-7. doi: 10.2144/04374ST03.
Measurement of transendothelial or transepithelial electrical resistances (TERs) is a straightforward in situ experimental approach to monitor the expression or modulation of barrier-forming cell-to-cell contacts (tight junctions) in cultured cells grown on porous filters. Although widely accepted, there is currently no device available to automatically measure the time course of TERs under ordinary cell culture conditions (37 degrees C, 5% or 10% CO2). This paper describes a development from our laboratory that is capable of following in parallel the TERs of several filter-grown cell layers with time and in an entirely computer-controlled fashion. The cell cultures can be followed even in long-term experiments without any manual assistance or opening of the incubator Besides reading TER values, this approach also returns the electrical capacitance of the cell layers, which is indicative of the expression of microvilli and other membrane extrusions. The device is based on reading the frequencydependent impedance of the cell layer, followed by equivalent circuit modeling to extract the cell-related parameters. It is compatible with several multi-well formats (up to 96 wells) and controlled by custom-designed software that reads, analyzes, and presents the data.
测量跨内皮或跨上皮电阻（TERs）是一种直接的原位实验方法，用于监测在多孔滤膜上生长的培养细胞中形成屏障的细胞间接触（紧密连接）的表达或调节。尽管该方法已被广泛接受，但目前尚无设备能够在普通细胞培养条件（37℃、5%或10%二氧化碳）下自动测量TERs的时间进程。本文介绍了我们实验室的一项成果，该成果能够以完全计算机控制的方式，随时间并行跟踪多个滤膜生长细胞层的TERs。即使在长期实验中，无需任何人工协助或打开培养箱，也能跟踪细胞培养情况。除了读取TER值外，该方法还能返回细胞层的电容，这可指示微绒毛和其他膜突出物的表达。该设备基于读取细胞层的频率依赖性阻抗，然后通过等效电路建模来提取与细胞相关的参数。它与多种多孔板格式（最多96孔）兼容，并由定制设计的软件控制，该软件可读取、分析和呈现数据。
