Epithelial Monolayer Integrity
Epithelial Monolayer Integrity - Related Videos
02:28
Assessing Transcytosis of E. coli Across Intestinal Epithelial Monolayers
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2025
Source: Islam, A., et al., Assessment of Intestinal Transcytosis of Neonatal Escherichia coli Bacteremia Isolates. J. Vis. Exp. (2023)This video demonstrates the procedure for assessing transcytosis of neonatal Escherichia coli across intestinal epithelial monolayers using transwell inserts, electrical resistance measurement, and bacterial quantification.
06:32
Development of Human Renal Tubular Epithelial Cell Primary Cultures in Monolayers and Three-Dimensional Conditions
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2025
Here, we describe the protocols for obtaining primary cultures of human renal tubular epithelial cells (HRTEC) from the kidney cortex to develop monolayers and three-dimensional cultures on an extracellular matrix (3D-HRTEC).
06:57
Generation of Murine Primary Colon Epithelial Monolayers from Intestinal Crypts
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Cited by 12 •
2021
In this protocol, we describe how to generate murine primary epithelial colon monolayers directly from intestinal crypts. We provide experimental approaches to generate confluent monolayers on permeable filters, confluent monolayers for scratch wound healing and biochemical studies, and sparse and confluent monolayers for immunofluorescence analysis.
11:48
Mouse- and Human-derived Primary Gastric Epithelial Monolayer Culture for the Study of Regeneration
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Cited by 9 •
2018
Here we describe a protocol for establishing and culturing human- and mouse-derived 3-dimensional (3D) gastric organoids, and the method for the transfer of 3D organoids to a 2-dimensional monolayer. The use of the gastric epithelial monolayer as a novel scratch-wound assay for regeneration studies is also described.
09:40
Organoid-Derived Epithelial Monolayer: A Clinically Relevant In Vitro Model for Intestinal Barrier Function
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Cited by 19 •
2021
Here, we describe the preparation of human organoid-derived intestinal epithelial monolayers for studying intestinal barrier function, permeability, and transport. As organoids represent original epithelial tissue response to external stimuli, these models combine the advantages of expandability of cell lines and the relevance and complexity of primary tissue.
FAQs
Tight junctions regulate movement through the space between neighboring cells, thereby controlling paracellular transport. Adherens junctions provide cell-to-cell adhesion that helps maintain tissue cohesion and organized cell positioning. Their coordinated activity allows the monolayer to remain continuous while preserving selective permeability, so disruption of either junctional system can alter barrier performance.
Apical-basal polarity organizes epithelial cells according to their tissue-facing and compartment-facing surfaces. This spatial organization supports the proper arrangement and function of junctional complexes, helping the monolayer separate tissue compartments. When polarity changes, junctional organization and selective transport may also change, providing a mechanistic link between cell organization and barrier integrity.
These stimuli can disturb cell adhesion, junctional organization, or the continuity of the monolayer. Such changes may modify paracellular transport and produce measurable alterations in permeability or electrical resistance. Examining the response to different stimulus types helps researchers determine whether a barrier has been disrupted and whether its organization can subsequently be restored.
Permeability measurements indicate how readily substances move across the epithelial barrier, whereas electrical resistance measurements provide a complementary assessment of barrier properties. Comparing these readouts with visible changes in junctional organization can distinguish functional alterations from structural ones. Together, they help characterize whether a stimulus compromises or restores epithelial barrier performance.
Assessment combines functional and structural observations rather than relying on a single readout. Researchers can measure permeability and electrical resistance, then examine changes in the organization of tight and adherens junctions. This combined approach connects transport behavior with cell architecture and helps evaluate how physical, chemical, or biological conditions affect the barrier.
Studying epithelial monolayer integrity is useful in research on barrier function, wound repair, inflammation, infection, and drug transport. In wound-repair studies, changes in continuity and junctional organization can indicate restoration. In inflammation or infection models, altered permeability and resistance can reveal barrier disruption, while transport studies use these measurements to evaluate passage across the epithelium.