AB079. SOH25_AB_131. Using the xCELLigence as a tool for real-time monitoring of cellular barrier integrity in vitro
Scientific Session

AB079. SOH25_AB_131. Using the xCELLigence as a tool for real-time monitoring of cellular barrier integrity in vitro

Laura O’Halloran1, Amira Mahdi2

1Faculty of Education & Health Services, School of Medicine, University of Limerick, Limerick, Ireland; 2Limerick Digital Cancer Research Centre, Health Research Institute, School of Medicine, University of Limerick, Limerick, Ireland

Background: Modelling cellular barriers is essential for understanding pathologies such as inflammatory bowel disease or blood-brain-barrier disruptions in brain metastasis. While in vitro models are cost-effective, they typically depend on endpoint transwell techniques lacking high throughput capabilities and limiting drug discovery. This study investigated the xCELLigence real-time cell analysis system, which measures changes in electrical impedance across cell barriers, as a dynamic and high throughput method for assessing cellular barrier integrity.

Methods: Caco-2 cells, derived from colon carcinoma, were seeded into xCELLigence E-16 plates, as differentiated monolayers. Barrier integrity was evaluated via electrical impedance. Ethanol treatment, a known barrier disrupter was used as a positive control. Barrier integrity was assessed following exposure to lipopolysaccharide (LPS) and breast cancer cell conditioned media over 4 days to simulate gut microbiota interactions and brain metastasis, respectively. xCELLigence system readouts were compared against traditional Transwell model end-point phenol red permeability assays. Results represent the average values from two biological replicates.

Results: Ethanol demonstrated concentration-dependent disruption of barrier integrity, with concentrations >5% significantly decreasing electrical impedance over 72 hours. LPS concentrations of 0.5 and 1 µg/mL caused a gradual but nonsignificant decline in electrical impedance at 96 hours. Conditioned breast cancer media demonstrated a decrease in electrical impedance 12 and 24 hours post-treatment. Findings from the xCELLigence system were consistent with results from traditional transwell assays for all tested conditions.

Conclusions: The xCELLigence system effectively detected real-time changes in barrier integrity offering a straightforward and scalable tool for modelling diverse biological barriers and supporting drug screening workflows.

Keywords: Blood-brain-barrier; Caco-2; gut barrier; real-time cell analysis; xCELLigence

Acknowledgments

None.

Footnote

Funding: None.

Conflicts of Interest: The authors have no conflicts of interest to declare.

Ethical Statement: The authors are accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.

Open Access Statement: This is an Open Access article distributed in accordance with the Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License (CC BY-NC-ND 4.0), which permits the non-commercial replication and distribution of the article with the strict proviso that no changes or edits are made and the original work is properly cited (including links to both the formal publication through the relevant DOI and the license). See: https://creativecommons.org/licenses/by-nc-nd/4.0/.

doi: 10.21037/map-25-ab079
Cite this abstract as: O’Halloran L, Mahdi A. AB079. SOH25_AB_131. Using the xCELLigence as a tool for real-time monitoring of cellular barrier integrity in vitro. Mesentery Peritoneum 2025;9:AB079.

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