On-chip epithelial barrier function assays using electrical impedance spectroscopy
On-chip epithelial barrier function assays using electrical impedance spectroscopy
A bio-impedance chip has been developed for real-time monitoring of the kinetics of epithelial cell monolayers in vitro. The human bronchial epithelial cell line (16-HBE 14o-) was cultured in Transwells(R) creating a sustainable and interactive model of the airway epithelium.
Conducting polymer polypyrrole (PPy) doped with polystyrene sulfonate (PSS) was electrochemically deposited onto the surface of gold-plated electrodes to reduce the influence of the electrical double layer on the impedance measurements. Finite element and equivalent circuit models were used to model and determine the electrical properties of the epithelial cell monolayer from the impedance spectra. Electrically tight, confluent monolayers of 16 HBE 14o- cells were treated with increasing concentrations of either Triton X-100 to solubilize cell membranes or ethylene glycol-bis(2-aminoethyl-ether)-N,N,N'N'-tetraacetic acid (EGTA) to disrupt cell-cell adhesion. Experimental impedance data showed that disruption of epithelial barrier function in response to Triton X-100 and EGTA can be successfully measured by the bio-impedance chip.
The results were consistent with the conventional hand-held trans-epithelial electrical resistance measurements. Immunofluorescent staining of the ZO-1 tight junction protein in the untreated and treated 16HBEs was performed to verify the disruption of the tight junctions by EGTA.
1611-1617
Sun, Tao
65fbe35a-ab15-4783-a5c3-54b2d05a9e38
Swindle, Emily J.
fe393c7a-a513-4de4-b02e-27369bd7e84f
Collins, Jane E.
be0e66f1-3036-47fa-9d7e-914c48710ba4
Holloway, Judith A.
f22f45f3-6fc8-4a4c-bc6c-24add507037c
Davies, Donna E.
7de8fdc7-3640-4e3a-aa91-d0e03f990c38
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174
21 June 2010
Sun, Tao
65fbe35a-ab15-4783-a5c3-54b2d05a9e38
Swindle, Emily J.
fe393c7a-a513-4de4-b02e-27369bd7e84f
Collins, Jane E.
be0e66f1-3036-47fa-9d7e-914c48710ba4
Holloway, Judith A.
f22f45f3-6fc8-4a4c-bc6c-24add507037c
Davies, Donna E.
7de8fdc7-3640-4e3a-aa91-d0e03f990c38
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174
Sun, Tao, Swindle, Emily J., Collins, Jane E., Holloway, Judith A., Davies, Donna E. and Morgan, Hywel
(2010)
On-chip epithelial barrier function assays using electrical impedance spectroscopy.
Lab on a Chip, 10 (12), .
(doi:10.1039/c000699h).
(PMID:20379587)
Abstract
A bio-impedance chip has been developed for real-time monitoring of the kinetics of epithelial cell monolayers in vitro. The human bronchial epithelial cell line (16-HBE 14o-) was cultured in Transwells(R) creating a sustainable and interactive model of the airway epithelium.
Conducting polymer polypyrrole (PPy) doped with polystyrene sulfonate (PSS) was electrochemically deposited onto the surface of gold-plated electrodes to reduce the influence of the electrical double layer on the impedance measurements. Finite element and equivalent circuit models were used to model and determine the electrical properties of the epithelial cell monolayer from the impedance spectra. Electrically tight, confluent monolayers of 16 HBE 14o- cells were treated with increasing concentrations of either Triton X-100 to solubilize cell membranes or ethylene glycol-bis(2-aminoethyl-ether)-N,N,N'N'-tetraacetic acid (EGTA) to disrupt cell-cell adhesion. Experimental impedance data showed that disruption of epithelial barrier function in response to Triton X-100 and EGTA can be successfully measured by the bio-impedance chip.
The results were consistent with the conventional hand-held trans-epithelial electrical resistance measurements. Immunofluorescent staining of the ZO-1 tight junction protein in the untreated and treated 16HBEs was performed to verify the disruption of the tight junctions by EGTA.
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Published date: 21 June 2010
Organisations:
Infection Inflammation & Immunity
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Local EPrints ID: 145569
URI: http://eprints.soton.ac.uk/id/eprint/145569
ISSN: 1473-0197
PURE UUID: f9b89a67-bb10-4574-b7ff-6a30e04ca87d
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Date deposited: 19 Apr 2010 10:55
Last modified: 14 Mar 2024 02:55
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Author:
Tao Sun
Author:
Hywel Morgan
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