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On-chip epithelial barrier function assays using electrical impedance spectroscopy

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.
1473-0197
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
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), 1611-1617. (doi:10.1039/c000699h). (PMID:20379587)

Record type: Article

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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More information

Published date: 21 June 2010
Organisations: Infection Inflammation & Immunity

Identifiers

Local EPrints ID: 145569
URI: https://eprints.soton.ac.uk/id/eprint/145569
ISSN: 1473-0197
PURE UUID: f9b89a67-bb10-4574-b7ff-6a30e04ca87d
ORCID for Emily J. Swindle: ORCID iD orcid.org/0000-0003-3644-7747
ORCID for Judith A. Holloway: ORCID iD orcid.org/0000-0002-2268-3071
ORCID for Donna E. Davies: ORCID iD orcid.org/0000-0002-5117-2991
ORCID for Hywel Morgan: ORCID iD orcid.org/0000-0003-4850-5676

Catalogue record

Date deposited: 19 Apr 2010 10:55
Last modified: 20 Jul 2019 01:28

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Contributors

Author: Tao Sun
Author: Jane E. Collins
Author: Donna E. Davies ORCID iD
Author: Hywel Morgan ORCID iD

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