Bischoff, Iris and Hornburger, Michael C. and Mayer, Bettina A. and Beyerle, Andrea and Wegener, Joachim and Fuerst, Robert (2016) Pitfalls in assessing microvascular endothelial barrier function: impedance-based devices versus the classic macromolecular tracer assay. SCIENTIFIC REPORTS, 6: 23671. ISSN 2045-2322,
Full text not available from this repository. (Request a copy)Abstract
The most frequently used parameters to describe the barrier properties of endothelial cells (ECs) in vitro are (i) the macromolecular permeability, indicating the flux of a macromolecular tracer across the endothelium, and (ii) electrical impedance of ECs grown on gold-film electrodes reporting on the cell layer's tightness for ion flow. Due to the experimental differences between these approaches, inconsistent observations have been described. Here, we present the first direct comparison of these assays applied to one single cell type (human microvascular ECs) under the same experimental conditions. The impact of different pharmacological tools (histamine, forskolin, Y-27632, blebbistatin, TRAP) on endothelial barrier function was analyzed by Transwell (R) tracer assays and two commercial impedance devices (xCELLigence (R), ECIS (R)). The two impedance techniques provided very similar results for all compounds, whereas macromolecular permeability readings were found to be partly inconsistent with impedance. Possible reasons for these discrepancies are discussed. We conclude that the complementary combination of both approaches is highly recommended to overcome the restrictions of each assay. Since the nature of the growth support may contribute to the observed differences, structure-function relationships should be based on cells that are consistently grown on either permeable or impermeable growth supports in all experiments.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | STRESS FIBER FORMATION; PROTEIN-KINASE; IN-VITRO; F-ACTIN; ZO-1 REDISTRIBUTION; CELL PERMEABILITY; MAMMALIAN-CELLS; THERMAL-INJURY; MYOSIN-II; PHOSPHORYLATION; |
| Subjects: | 500 Science > 540 Chemistry & allied sciences |
| Divisions: | Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik > Bioanalytik und Biosensorik (Prof. Joachim Wegener) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 26 Mar 2019 06:11 |
| Last Modified: | 26 Mar 2019 06:11 |
| URI: | https://pred.uni-regensburg.de/id/eprint/3219 |
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