Hajek, Kathrin and Wegener, Joachim (2017) Independent impedimetric analysis of two cell populations co-cultured on opposite sides of a porous support. EXPERIMENTAL CELL RESEARCH, 351 (1). pp. 121-126. ISSN 0014-4827, 1090-2422
Full text not available from this repository. (Request a copy)Abstract
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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | CAPILLARY ENDOTHELIAL-CELLS; PLEXUS EPITHELIAL-CELLS; BLOOD-BRAIN-BARRIER; IN-VITRO; TIGHT JUNCTIONS; TRANSPORT; DYNAMICS; CULTURE; MODEL; Impedance analysis; Transepithelial electrical resistance (TEER); Co-cultures; MDCK; Label-free; Cell-based assays; Barrier function |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik > Bioanalytik und Biosensorik (Prof. Joachim Wegener) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 14 Dec 2018 13:01 |
| Last Modified: | 14 Feb 2019 09:19 |
| URI: | https://pred.uni-regensburg.de/id/eprint/439 |
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