Functional optimization of electric cell-substrate impedance sensing (ECIS) using human corneal epithelial cells

Abstract

An intact epithelium is key to maintaining corneal integrity and barrier function which can lead to impaired ocular defense and sight-threatening opacity when compromised. Electrical cell-substrate impedance sensing or ECIS is a non-invasive method to measure real-time cellular behaviors including barrier function and cell migration. The current study uses ECIS technology to assess and optimize human telomerase-immortalized corneal epithelial cells (HUCLs) to generate quantifiable measurements that accurately reflect changes in cell behavior in vitro. Five cell densities were assessed in two different media to determine the optimal conditions for monitoring of cellular behavior over time. Parameters of evaluation included: overall impedance (Z), barrier resistance (R), cell capacitance (C), and mathematical modeling of the R data to further generate Rb (the electrical resistance between HUCLs), α (the resistance between the HUCLs and the substrate), and Cm (the capacitance of the cell membrane) measurements. All parameters of assessment strongly indicated DMEM/F12 at 60,000 cells as the optimal condition for ECIS assessment of HUCLs. Furthermore, this work highlights the ability of the sensitive ECIS biosensor technology to comprehensively and quantitatively assess corneal epithelial cell structure and function and the importance of optimizing not only cell density, but choice of media used for in vitro culturing.

Cell-Based Electrical Impedance Platform to Evaluate Zika Virus Inhibitors in Real Time

“…The use of the CEI technology is limited to adherent cell types, as the assay's principle relies on the properties of adherent cells to spread across the electrode-embedded wells. Well surface coatings such as fibronectin can be used to improve cell attachment 29 . The methodology has some other drawbacks, the first related to its cost.…”

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“…The use of the CEI technology is limited to adherent cell types, as the assay's principle relies on the properties of adherent cells to spread across the electrode-embedded wells. Well surface coatings such as fibronectin can be used to improve cell attachment 29 . The methodology has some other drawbacks, the first related to its cost.…”