A simple approach to measurement of electrical parameters of cultured epithelial monolayers: use in assessing neutrophil-epithelial interactions

JL Madara, S Colgan, A Nusrat, C Delp… - Journal of tissue culture …, 1992 - Springer
Journal of tissue culture methods, 1992Springer
Typically, electrophysiologic studies of epithelial monolayers are either performed in
formalized Ussing chamber systems which yield highly accurate results or in simple setups
using recording devices which have limitations in accuracy or the range of measurements
which can be performed or both. Here we detail a simple method of interfacing traditionally
accurate Ussing chambers systems with commercially available filter supports on which
epithelial monolayers can be grown. We also detail simple methods for growing inverted …
Summary
Typically, electrophysiologic studies of epithelial monolayers are either performed in formalized Ussing chamber systems which yield highly accurate results or in simple setups using recording devices which have limitations in accuracy or the range of measurements which can be performed or both. Here we detail a simple method of interfacing traditionally accurate Ussing chambers systems with commercially available filter supports on which epithelial monolayers can be grown. We also detail simple methods for growing inverted monolayers, suitable for electrophysiologic assays, for use in studies where cells or particles must be layered by gravity on the undersurface (basolateral pole) of the filter. Both approaches allow experiments to be performed on large numbers of monolayers synchronously. As an example of the use of this system, we analyze the sequelae of neutrophil migration across monolayers of the intestinal cell line T84. Neutrophil migration across monolayers can occur in either direction, is dependent on neutrophil surface β2 integrins, and is paralleled by a decrease in epithelial barrier function as detected electrically. We have previously shown in formal Ussing chamber studies that neutrophil-epithelial interactions elicit a modest short-circuit current indicative of electrogenic ion transport. We show here that this short-circuit current response can be readily detected using the simple approach described.
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