Cell Barrier Function
Endothelial and epithelial cells form vital barriers that regulate the movement of molecules and ions between tissues. When these barriers are disrupted—by inflammation, disease, or compound exposure—cell health and function decline. Traditional in-vitro methods such as solute permeability and TEER assays are labor-intensive, low-throughput, and offer only limited time-point data.
The xCELLigence RTCA System provides a label-free, real-time, impedance-based alternative that continuously monitors barrier integrity under physiological conditions. Because the readout is noninvasive, users can combine RTCA measurements with microscopy and other orthogonal assays on the same cells—eliminating the need for transwell-based workflows.
Limitations of Traditional Assays
Solute Permeability Assays
- No information on cell health or confluency
- Risk of leakage from uneven monolayers
- Cannot track barrier recovery
- Costly and low throughput
Classical TEER Assays
- No visibility into cell quality or confluency
- Susceptible to leakage and variability
- Labor-intensive, contamination risk
- Only snapshots at discrete time points
Key Benefits of xCELLigence RTCA
Direct, label-free, continuous monitoring.
Real-time measurement of both barrier disruption and recovery.
Noninvasive readout enables complementary microscopy.
TEER-like assessment directly on Agilent E-Plates - no transwells needed.
xCELLigence TEER assay – a Plate Assay
A label-free, real-time alternative to solute permeability and transendothelial electrical resistance (TEER) assays, monitoring of cell barrier function disruption and recovery in extreme simplicity:
STEP 1: Seed Cells, let the Cell Index grow to plateau
STEP 2: Add treatment, observed barrier function disruption and recovery
Data Comparison from Solute Permeability Assay and xCELLigence TEER assay
The protective effect of Activated Protein C (APC) on endothelial barrier function assessed by solute permeability assays and RTCA platform. EA.hy926 cells were pre-incubated with APC, and the cell barrier dysfunction induced by thrombin was assessed by measuring (A) Evans blue permeability and (B) FITC permeability. (C) Dynamic impedance-based monitoring of thrombin induced endothelial barrier dysfunction. (D) The maximal change in the endothelial barrier function as percentage of the control derived from Cell Index changes.
Data and figures courtesy of F. Stavenuiter, E. Bouwens, R. Sinha, L. Mosnier, and J. H. Griffin, Dept. of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA.
Realted Publications
- VEGF receptor 2 inhibitor nintedanib completely reverts VEGF-A165-induced disturbances of barriers formed by retinal endothelial cells or long-term cultivated ARPE-19 cells. Deissler HL, Stutzer JN, Lang GK, Grisanti S, Lang GE, Ranjbar M. Exp Eye Res. 2020 Mar 14;194:108004.
- Single Triglyceride-Rich Meal Destabilizes Barrier Functions and Initiates Inflammatory Processes of Endothelial Cells.Gorzelak-Pabiś P, Wozniak E, Wojdan K, Chalubinski M, Broncel M.J Interferon Cytokine Res. 2020 Jan;40(1):43-53.
- Culture Model for Non-human Primate Choroid Plexus.Delery EC, MacLean AG.Front Cell Neurosci. 2019 Aug 28;13:396.
- The functional and inflammatory response of brain endothelial cells to Toll-Like Receptor agonists. Johnson RH, Kho DT, O' Carroll SJ, Angel CE, Graham ES. Sci Rep. 2018 Jul 4;8(1):10102.
- CD146 coordinates brain endothelial cell-pericyte communication for blood-brain barrier development. Chen J, Luo Y, Hui H, Cai T, Huang H, Yang F, Feng J, Zhang J, Yan X. Proc Natl Acad Sci U S A. 2017 Sep 5;114(36):E7622-E7631
- Pitfalls in assessing microvascular endothelial barrier function: impedance-based devices versus the classic macromolecular tracer assay. Bischoff I, Hornburger MC, Mayer BA, Beyerle A, Wegener J, Fürst R. Sci Rep. 2016 Mar 30;6:23671
- Apolipoprotein E Receptor 2 Mediates Activated Protein C–Induced Endothelial Akt Activation and Endothelial Barrier Stabilization. Sinha RK, Yang XV, Fernández JA, Xu X, Mosnier LO, Griffin JH. Arterioscler Thromb Vasc Biol. 2016 Mar;36(3):518-24.
- CCM1–ICAP-1 complex controls β1 integrin–dependent endothelial contractility and fibronectin remodeling. Faurobert E, Rome C, Lisowska J, Manet-Dupé S, Boulday G, Malbouyres M, Balland M, Bouin AP,Kéramidas M, Bouvard D, Coll JL, Ruggiero F, Tournier-Lasserve E, Albiges-Rizo C. J Cell Biol. 2013 Aug 5;202(3):545-61.
- Vinculin-dependent Cadherin mechanosensing regulates efficient epithelial barrier formation. Twiss F, Le Duc Q, Van Der Horst S, Tabdili H, Van Der Krogt G, Wang N, Rehmann H, Huveneers S,Leckband DE, De Rooij J. Biology Open. 2012; doi:10.1242/bio.20122428
- An inverted blood-brain barrier model that permits interactions between glia and inflammatory stimuli. Sansing HA, Renner NA, MacLean AG. Journal of neuroscience methods. 2012;207(1):91–6.
- A dynamic real-time method for monitoring epithelial barrier function in vitro. Sun M, Fu H, Cheng H, Cao Q, Zhao Y, Mou X, Zhang X, Liu X, Ke Y. Analytical biochemistry. 2012;425(2):96–103.