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.
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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August 2017
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, 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), .
(doi:10.1016/j.tice.2017.06.003).
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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Ratnayaka_et_al_manuscript_2017
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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
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Local EPrints ID: 411709
URI: http://eprints.soton.ac.uk/id/eprint/411709
ISSN: 0040-8166
PURE UUID: 211513f5-656e-4c07-9b07-47ca09390718
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Date deposited: 01 Feb 2018 17:33
Last modified: 16 Mar 2024 05:28
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Author:
Savannah Lynn
Author:
Gareth Ward
Author:
Eloise Keeling
Author:
Jenny Scott
Author:
David A. Johnston
Author:
Anton Page
Author:
Enrique Cuan Urquizo
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