Evaluating Epithelial Barrier Function

Transepithelial electrical resistance (TEER) of Caco-2 cells was measured before and after the exposure to the bacteria using the Millicell ERS system (Millipore, Billerica, MA). Furthermore, we analyzed the epithelial tight junction integrity as per Koo et al. [21 (link)]. After exposure to L. monocytogenes, the tight junction permeability was analyzed using Dextran^FITC (Mr 3–5 kDa; Sigma) permeability through the transwell filter inserts. Fluorescence of the samples collected from the apical and basolateral chambers was read in a SpectraMax Gemini EM fluorescent plate reader (Molecular Devices; Sunnyvale, CA).

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Mathipa M.G., Bhunia A.K, & Thantsha M.S. (2019). Internalin AB-expressing recombinant Lactobacillus casei protects Caco-2 cells from Listeria monocytogenes-induced damages under simulated intestinal conditions. PLoS ONE, 14(7), e0220321.

Publication 2019

Millicell ERS system DextranFITC SpectraMax Gemini EM

Bacteria Caco 2 cells Devices Electrical resistance Fluorescence Permeability Tight junction

Corresponding Organization : University of Pretoria

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Variable analysis

independent variables

Exposure to L. monocytogenes

dependent variables

Transepithelial electrical resistance (TEER) of Caco-2 cells
Epithelial tight junction integrity
Dextran^FITC (Mr 3–5 kDa) permeability through the transwell filter inserts

control variables

Caco-2 cells
Millicell ERS system (Millipore, Billerica, MA)
Dextran^FITC (Mr 3–5 kDa; Sigma)
SpectraMax Gemini EM fluorescent plate reader (Molecular Devices; Sunnyvale, CA)

controls

Positive control: Not specified
Negative control: Not specified

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