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Functional characterization of a novel 3D model of the epithelial-mesenchymal trophic unit

Functional characterization of a novel 3D model of the epithelial-mesenchymal trophic unit
Functional characterization of a novel 3D model of the epithelial-mesenchymal trophic unit
Background/Aim: Epithelial-mesenchymal communication plays a key role in tissue homeostasis and abnormal signaling contributes to chronic airways disease such as COPD. Most in vitro models are limited in complexity and poorly represent this epithelial-mesenchymal trophic unit. We postulated that cellular outgrowth from bronchial tissue would enable development of a mucosal structure that recapitulates better in vivo tissue architecture.

Materials and Methods: Bronchial tissue was embedded in Matrigel and outgrowth cultures monitored using time-lapse microscopy, electrical resistance, light and electron microscopy. Cultures were challenged repetitively with cigarette smoke extract (CSE).

Results: The outgrowths formed as a multicellular sheet with motile cilia becoming evident as the Matrigel was remodeled to provide an air interface; cultures were viable for more than one year. Immunofluorescence and electron microscopy (EM) identified an upper layer of mucociliary epithelium and a lower layer of highly organized extracellular matrix (ECM) interspersed with fibroblastic cells separated by a basement membrane. EM analysis of the mucosal construct after repetitive exposure of to CSE revealed epithelial damage, loss of cilia, and ECM remodeling, as occurs in vivo.

Conclusions: We have developed a robust bronchial mucosal model. The structural changes observed following CSE exposure suggest the model should have utility for drug discovery and preclinical testing, especially those targeting airway remodeling.
0190-2148
82-92
Bucchieri, Fabio
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Pitruzzella, Alessandro
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Fucarino, Alberto
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Marino Gammazza, Antonella
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Bavisotto, Celeste
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Marciano, Vito
703c45c4-b614-41f8-bc6f-122268be0df5
Cajozzo, Massimo
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Lo Iacono, Giorgio
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Marchese, Roberto
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Zummo, Giovanni
54897ebe-00d6-47b9-9914-fb350deb7b14
Holgate, Stephen T.
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Davies, Donna E.
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Bucchieri, Fabio
830cf621-3e39-42ab-b0ea-7d5dc43bf032
Pitruzzella, Alessandro
378be3bd-b7fc-4615-bd90-7a3753077a46
Fucarino, Alberto
95c722bb-5f4d-43b7-a173-c72e40b3c792
Marino Gammazza, Antonella
b8683acd-7d10-4e5a-9224-ff7b283d7a65
Bavisotto, Celeste
717dcc26-8bec-440a-ac24-0c6aa6283411
Marciano, Vito
703c45c4-b614-41f8-bc6f-122268be0df5
Cajozzo, Massimo
44bf71df-1506-43d0-82ca-cadcee0db4d1
Lo Iacono, Giorgio
4a69485f-c403-4ebc-985f-442682d01456
Marchese, Roberto
b4d3b606-24d5-43cc-ae8d-f9a43fd548ee
Zummo, Giovanni
54897ebe-00d6-47b9-9914-fb350deb7b14
Holgate, Stephen T.
2e7c17a9-6796-436e-8772-1fe6d2ac5edc
Davies, Donna E.
7de8fdc7-3640-4e3a-aa91-d0e03f990c38

Bucchieri, Fabio, Pitruzzella, Alessandro, Fucarino, Alberto, Marino Gammazza, Antonella, Bavisotto, Celeste, Marciano, Vito, Cajozzo, Massimo, Lo Iacono, Giorgio, Marchese, Roberto, Zummo, Giovanni, Holgate, Stephen T. and Davies, Donna E. (2017) Functional characterization of a novel 3D model of the epithelial-mesenchymal trophic unit. Experimental Lung Research, 43 (2), 82-92. (doi:10.1080/01902148.2017.1303098).

Record type: Article

Abstract

Background/Aim: Epithelial-mesenchymal communication plays a key role in tissue homeostasis and abnormal signaling contributes to chronic airways disease such as COPD. Most in vitro models are limited in complexity and poorly represent this epithelial-mesenchymal trophic unit. We postulated that cellular outgrowth from bronchial tissue would enable development of a mucosal structure that recapitulates better in vivo tissue architecture.

Materials and Methods: Bronchial tissue was embedded in Matrigel and outgrowth cultures monitored using time-lapse microscopy, electrical resistance, light and electron microscopy. Cultures were challenged repetitively with cigarette smoke extract (CSE).

Results: The outgrowths formed as a multicellular sheet with motile cilia becoming evident as the Matrigel was remodeled to provide an air interface; cultures were viable for more than one year. Immunofluorescence and electron microscopy (EM) identified an upper layer of mucociliary epithelium and a lower layer of highly organized extracellular matrix (ECM) interspersed with fibroblastic cells separated by a basement membrane. EM analysis of the mucosal construct after repetitive exposure of to CSE revealed epithelial damage, loss of cilia, and ECM remodeling, as occurs in vivo.

Conclusions: We have developed a robust bronchial mucosal model. The structural changes observed following CSE exposure suggest the model should have utility for drug discovery and preclinical testing, especially those targeting airway remodeling.

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Bucchieri et al_accepted MS and figs - Accepted Manuscript
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Accepted/In Press date: 2 March 2017
e-pub ahead of print date: 3 April 2017
Published date: 2017
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 407571
URI: https://eprints.soton.ac.uk/id/eprint/407571
ISSN: 0190-2148
PURE UUID: d11b303b-ca95-4491-9653-6f37adffa52b
ORCID for Donna E. Davies: ORCID iD orcid.org/0000-0002-5117-2991

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Date deposited: 13 Apr 2017 01:10
Last modified: 10 Dec 2019 06:09

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Contributors

Author: Fabio Bucchieri
Author: Alessandro Pitruzzella
Author: Alberto Fucarino
Author: Antonella Marino Gammazza
Author: Celeste Bavisotto
Author: Vito Marciano
Author: Massimo Cajozzo
Author: Giorgio Lo Iacono
Author: Roberto Marchese
Author: Giovanni Zummo
Author: Donna E. Davies ORCID iD

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