Wegener J, Sieber M, Galla H J
Institut für Biochemie, Westfälische Wilheims-Universität Münster, Germany.
J Biochem Biophys Methods. 1996 Jul 10;32(3):151-70. doi: 10.1016/0165-022x(96)00005-x.
The present study describes a new method to determine transepithelial and transendothelial electrical resistances (TER) of cultured cell monolayers which is based on impedance analysis. To obtain impedance data of the epithelia or endothelia under investigation, we developed special measuring chambers that allow to culture the cells on gold surfaces that are used as measuring electrodes. Impedance analysis is carried out in the frequency range from 1 to 10(5) s-1 under normal culture conditions using a self-developed continuous wave impedance spectrometer. Evaluation of impedance data is achieved by fitting (NLSQ) the parameters of appropriate equivalent circuits to the experimental data. We investigated cell monolayers of primary cultured endothelial cells isolated from porcine brain microvessels, epithelial cells from porcine choroid plexus as well as those of the epithelial cell line MDCK. Transepithelial resistances were found to be in good agreement with published data. The main advantages of the new technique are the ability (i) to use multi-electrode arrays that allow to determine TERs at different locations of a given cell monolayer; (ii) to carry out impedance analysis under normal culture conditions; and (iii) to obtain TER values of cell monolayers grown on impermeable supports, which means that conditions cells are normally exposed to in ordinary culture dishes are maintained.
本研究描述了一种基于阻抗分析来测定培养细胞单层的跨上皮和跨内皮电阻（TER）的新方法。为了获取被研究上皮或内皮的阻抗数据，我们开发了特殊的测量室，其允许在用作测量电极的金表面上培养细胞。使用自行研制的连续波阻抗光谱仪，在正常培养条件下，于1至10⁵ s⁻¹ 的频率范围内进行阻抗分析。通过将合适的等效电路参数拟合（非线性最小二乘法）到实验数据来实现阻抗数据的评估。我们研究了从猪脑微血管分离的原代培养内皮细胞、猪脉络丛上皮细胞以及上皮细胞系MDCK的细胞单层。发现跨上皮电阻与已发表的数据高度一致。这项新技术的主要优点在于：（i）能够使用多电极阵列，从而可以在给定细胞单层的不同位置测定TER；（ii）能够在正常培养条件下进行阻抗分析；（iii）能够获得在不可渗透支持物上生长的细胞单层的TER值，这意味着维持了细胞在普通培养皿中通常所接触的条件。
