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The cells have >90% expression of CD105 (endoglin), CD31 (PECAM-1) and CD144 (VE-cad) as determined by flow cytometry analysis.
(A) When immunostained for von Willebrand Factor (vWF), the cells showed characteristic Weibel-Palade body staining. (B) In a thick layer of Matrigel, the cells exhibited the capacity to form tubes. (C) To assess the barrier function, the cells were immunostained for the tight junction protein ZO-1.
iCell Endothelial Cells exhibit barrier function activity that can be disrupted by treatment with thrombin and reliably assessed using impedance-based platforms. iCell Endothelial Cells provide an in vitro test system that recapitulates native human endothelium properties and functions while the xCELLigence RTCA Cardio System provides a label-free technology for non-invasive monitoring of cell behavior and viability. The methods and results presented here highlight how to gather relevant data on human endothelial viability and barrier function. Together, iCell Endothelial Cells and impedance-based technologies offer a valuable cell model system for understanding the endothelial barrier characteristics, mechanisms of endothelial barrier dysfunction, and dynamic modulation of the endothelium permeability, enabling a wide range of applications in academic and pharmaceutical research. iCell Endothelial Cells were cultured for 48 hours on the E-Plate in Complete iCell Endothelial Cells Maintenance Medium before the addition of thrombin. (A) The representative Cell Index curves showed the dose-dependent disruption effect of thrombin on the barrier function and the recovery phase. (B) The percentage of Cell Index compared to pre-treatment baseline levels was calculated at the time of thrombin addition, at 9 hours post-treatment, or at 21 hours post-treatment (mean ± SD, n = 2 wells).
iCell Endothelial Cells are amenable to a variety of assays including: