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

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 b...

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Autores principales: Ebrahim, Abdul Shukkur, Ebrahim, Thanzeela, Kani, Hussein, Ibrahim, Ahmed S., Carion, Thomas W., Berger, Elizabeth A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9391335/
https://www.ncbi.nlm.nih.gov/pubmed/35986158
http://dx.doi.org/10.1038/s41598-022-18182-z
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author Ebrahim, Abdul Shukkur
Ebrahim, Thanzeela
Kani, Hussein
Ibrahim, Ahmed S.
Carion, Thomas W.
Berger, Elizabeth A.
author_facet Ebrahim, Abdul Shukkur
Ebrahim, Thanzeela
Kani, Hussein
Ibrahim, Ahmed S.
Carion, Thomas W.
Berger, Elizabeth A.
author_sort Ebrahim, Abdul Shukkur
collection PubMed
description 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 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 R(b) (the electrical resistance between HUCLs), α (the resistance between the HUCLs and the substrate), and C(m) (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.
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spelling pubmed-93913352022-08-21 Functional optimization of electric cell-substrate impedance sensing (ECIS) using human corneal epithelial cells Ebrahim, Abdul Shukkur Ebrahim, Thanzeela Kani, Hussein Ibrahim, Ahmed S. Carion, Thomas W. Berger, Elizabeth A. Sci Rep Article 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 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 R(b) (the electrical resistance between HUCLs), α (the resistance between the HUCLs and the substrate), and C(m) (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. Nature Publishing Group UK 2022-08-19 /pmc/articles/PMC9391335/ /pubmed/35986158 http://dx.doi.org/10.1038/s41598-022-18182-z Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Ebrahim, Abdul Shukkur
Ebrahim, Thanzeela
Kani, Hussein
Ibrahim, Ahmed S.
Carion, Thomas W.
Berger, Elizabeth A.
Functional optimization of electric cell-substrate impedance sensing (ECIS) using human corneal epithelial cells
title Functional optimization of electric cell-substrate impedance sensing (ECIS) using human corneal epithelial cells
title_full Functional optimization of electric cell-substrate impedance sensing (ECIS) using human corneal epithelial cells
title_fullStr Functional optimization of electric cell-substrate impedance sensing (ECIS) using human corneal epithelial cells
title_full_unstemmed Functional optimization of electric cell-substrate impedance sensing (ECIS) using human corneal epithelial cells
title_short Functional optimization of electric cell-substrate impedance sensing (ECIS) using human corneal epithelial cells
title_sort functional optimization of electric cell-substrate impedance sensing (ecis) using human corneal epithelial cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9391335/
https://www.ncbi.nlm.nih.gov/pubmed/35986158
http://dx.doi.org/10.1038/s41598-022-18182-z
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