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Review
. 2022 Feb 9;5(1):17.
doi: 10.3390/mps5010017.

In Vitro Methods for Measuring the Permeability of Cell Monolayers

Radoslaw Bednarek  1
Affiliations
Review

In Vitro Methods for Measuring the Permeability of Cell Monolayers

Radoslaw Bednarek. Methods Protoc. .

Abstract

Cell monolayers, including endothelial and epithelial cells, play crucial roles in regulating the transport of biomolecules to underlying tissues and structures via intercellular junctions. Moreover, the monolayers form a semipermeable barrier across which leukocyte transmigration is tightly regulated. The inflammatory cytokines can disrupt the epithelial and endothelial permeability, thus the reduced barrier integrity is a hallmark of epithelial and endothelial dysfunction related with numerous pathological conditions, including cancer-related inflammation. Therefore, the assessment of barrier function is critical in in vitro models of barrier-forming tissues. This review summarizes the commercially available in vitro systems used to measure the permeability of cellular monolayers. The presented techniques are separated in two large groups: macromolecular tracer flux assays, and electrical impedance measurement-based permeability assays. The presented techniques are briefly described and compared.

Keywords: dysfunction; endothelium; epithelium; fluorescent tracer; microfluidics; permeability; transendothelial resistance.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
The overview of a transwell support system (taken from https://tumomics.creative-proteomics.com/, accessed on 28 June 2021, with permission).
Figure 2
Figure 2
The overview of EVOM3 (epithelial volt–ohm meter) system with all its components, used for the transepithelial/transendothelial electrical resistance (TEER) measurements (from left to right: A/C power supply and charger, STX2-Plus electrode, EVOM3 meter and the optional Endohm-6G chamber electrode). (A) the general view of the whole EVOM3 system. (B) The EVOM3 meter with the 1 kOhm test resistor inserted into the electrode port: the display shows the actual resistance of the test resistor. (C) The body of the STX2-Plus chopstick electrode. (D,E) The proper adjustment of STX2-Plus electrode to fit the insert with a 6.5 mm diameter (compatible with 24-well plates): the outer electrode reaches close to the bottom of the well plate, while the inner electrode is close to the membrane inside the insert, without touching it. (F) The Endohm-6G chamber electrode with the 6.5 mm cell culture insert inside the chamber, connected with the EVOM3 meter by the Endohm cable. (G) The detailed view of the Endohm-6G chamber with the cell culture insert.
Figure 3
Figure 3
Schematic drawing of a circulating Ussing chamber (taken from [30], license no. 5112401256947).
See this image and copyright information in PMC

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