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Ex-vivo models of the Retinal Pigment Epithelium (RPE) in long-term culture faithfully recapitulate key structural and physiological features of native RPE

Ex-vivo models of the Retinal Pigment Epithelium (RPE) in long-term culture faithfully recapitulate key structural and physiological features of native RPE
Ex-vivo models of the Retinal Pigment Epithelium (RPE) in long-term culture faithfully recapitulate key structural and physiological features of native RPE
The Retinal Pigment Epithelium (RPE) forms the primary site of pathology in several blinding retinopathies. RPE cultures are being continuously refined so that dynamic disease processes in this important monolayer can be faithfully studied outside the eye over longer periods. The RPE substrate, which mimics the supportive Bruch’s membrane (BrM), plays a key role in determining how well in-vitro cultures recapitulate native RPE cells. Here, we evaluate how two different types of BrM substrates; (1) a commercially-available polyester transwell membrane, and (2) a novel electrospun scaffold developed in our laboratory, could support the generation of realistic RPE tissues in culture. Our findings reveal that both substrates were capable of supporting long-lasting RPE monolayers with structural and functional specialisations of in-situ RPE cells. These cultures were used to study autofluorescence and barrier formation, as well as activities such as outer-segment internalisation/trafficking and directional secretion of key proteins; the impairment of which underlies retinal disease. Hence, both substrates fulfilled important criteria for generating authentic in-vitro cultures and act as powerful tools to study RPE pathophysiology. However, RPE grown on electrospun scaffolds may be better suited to studying complex RPE-BrM interactions such as the formation of drusen-like deposits associated with early retinal disease.
0040-8166
447-460
Lynn, Savannah
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Ward, Gareth
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Keeling, Eloise
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Scott, Jenny
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Cree, Angela J.
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Johnston, David A.
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Page, Anton
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Cuan Urquizo, Enrique
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Bhaskar, Atul
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Grossel, Martin C.
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Tumbarello, David A.
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Newman, Tracey A.
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Lotery, Andrew J.
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Ratnayaka, J. Arjuna
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Lynn, Savannah
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Ward, Gareth
250b5aa5-a94a-49ec-9569-9250d8e2607b
Keeling, Eloise
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Scott, Jenny
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Cree, Angela J.
6724b71b-8828-4abb-971f-0856c2af555e
Johnston, David A.
b41163c9-b9d2-425c-af99-2a357204014e
Page, Anton
76ebbfb8-4fe3-495c-afff-1f2f34977fee
Cuan Urquizo, Enrique
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Bhaskar, Atul
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Grossel, Martin C.
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Tumbarello, David A.
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Newman, Tracey A.
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Lotery, Andrew J.
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Ratnayaka, J. Arjuna
002499b8-1a9f-45b6-9539-5ac145799dfd

Lynn, Savannah, Ward, Gareth, Keeling, Eloise, Scott, Jenny, Cree, Angela J., Johnston, David A., Page, Anton, Cuan Urquizo, Enrique, Bhaskar, Atul, Grossel, Martin C., Tumbarello, David A., Newman, Tracey A., Lotery, Andrew J. and Ratnayaka, J. Arjuna (2017) Ex-vivo models of the Retinal Pigment Epithelium (RPE) in long-term culture faithfully recapitulate key structural and physiological features of native RPE. Tissue and Cell, 49 (4), 447-460. (doi:10.1016/j.tice.2017.06.003).

Record type: Article

Abstract

The Retinal Pigment Epithelium (RPE) forms the primary site of pathology in several blinding retinopathies. RPE cultures are being continuously refined so that dynamic disease processes in this important monolayer can be faithfully studied outside the eye over longer periods. The RPE substrate, which mimics the supportive Bruch’s membrane (BrM), plays a key role in determining how well in-vitro cultures recapitulate native RPE cells. Here, we evaluate how two different types of BrM substrates; (1) a commercially-available polyester transwell membrane, and (2) a novel electrospun scaffold developed in our laboratory, could support the generation of realistic RPE tissues in culture. Our findings reveal that both substrates were capable of supporting long-lasting RPE monolayers with structural and functional specialisations of in-situ RPE cells. These cultures were used to study autofluorescence and barrier formation, as well as activities such as outer-segment internalisation/trafficking and directional secretion of key proteins; the impairment of which underlies retinal disease. Hence, both substrates fulfilled important criteria for generating authentic in-vitro cultures and act as powerful tools to study RPE pathophysiology. However, RPE grown on electrospun scaffolds may be better suited to studying complex RPE-BrM interactions such as the formation of drusen-like deposits associated with early retinal disease.

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Accepted/In Press date: 16 June 2017
e-pub ahead of print date: 19 June 2017
Published date: August 2017
Organisations: Clinical Neurosciences, Computational Engineering & Design Group, Academic, Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 411709
URI: https://eprints.soton.ac.uk/id/eprint/411709
ISSN: 0040-8166
PURE UUID: 211513f5-656e-4c07-9b07-47ca09390718
ORCID for Savannah Lynn: ORCID iD orcid.org/0000-0003-2513-3144
ORCID for David A. Johnston: ORCID iD orcid.org/0000-0001-6703-6014
ORCID for Martin C. Grossel: ORCID iD orcid.org/0000-0001-7469-6854
ORCID for David A. Tumbarello: ORCID iD orcid.org/0000-0002-5169-0561
ORCID for Tracey A. Newman: ORCID iD orcid.org/0000-0002-3727-9258
ORCID for Andrew J. Lotery: ORCID iD orcid.org/0000-0001-5541-4305
ORCID for J. Arjuna Ratnayaka: ORCID iD orcid.org/0000-0002-1027-6938

Catalogue record

Date deposited: 01 Feb 2018 17:33
Last modified: 03 Dec 2019 06:01

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Contributors

Author: Savannah Lynn ORCID iD
Author: Gareth Ward
Author: Eloise Keeling
Author: Jenny Scott
Author: Angela J. Cree
Author: David A. Johnston ORCID iD
Author: Anton Page
Author: Enrique Cuan Urquizo
Author: Atul Bhaskar

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