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Development and characterization of an in vitro system of the human retina using cultured cell lines / Rachel, Churm; Sarah, Prior; David, Owens; Becky, Thomas

Clinical & Experimental Ophthalmology

Swansea University Authors: Rachel, Churm, Sarah, Prior, David, Owens, Becky, Thomas

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DOI (Published version): 10.1111/ceo.13578

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Background: Previously developed in vitro cultures of the human retina have been solo or dual cell cultures. We developed a triple-cell culture in vitro model utilizing a membrane system to produce a better representation of a functional and morphological human retina. Methods: Retinal microvascular...

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Published in: Clinical & Experimental Ophthalmology
ISSN: 1442-6404 1442-9071
Published: 2019
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URI: https://cronfa.swan.ac.uk/Record/cronfa50970
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spelling 2019-08-15T13:19:07.7882121 v2 50970 2019-07-01 Development and characterization of an in vitro system of the human retina using cultured cell lines c6cd8267ff0b13f2ea333bbfefdae144 0000-0001-9855-6282 Rachel Churm Rachel Churm true false cdda101035997acfaa6fdf17097f52b2 0000-0001-8703-8092 Sarah Prior Sarah Prior true false 2fd4b7c3f82c6d3bd546eff61ff944e9 0000-0003-1002-1238 David Owens David Owens true false e83b45ec71428bd748ce201048f43d6a Becky Thomas Becky Thomas true false 2019-07-01 STSC Background: Previously developed in vitro cultures of the human retina have been solo or dual cell cultures. We developed a triple-cell culture in vitro model utilizing a membrane system to produce a better representation of a functional and morphological human retina. Methods: Retinal microvascular endothelial cells (HRMVEC/ACBRI181, Cell systems), retinal pigment epithelium cells (RPE/ARPE-19, ATCC) and Müller glial cells (MIO-M1, UCL) were grown in a triple-culture. Our optimized triple-culture media contained a mix of specific endothelial medium and high glucose Dulbecco's Modified Eagle's medium (DMEM), where all three layers were viable for up to 5 days. Co-culture effect on morphological changes (cell staining) and gene expression of functional genes (pigment epithelial derived factor (PEDF) and vascular endothelial growth factor (VEGF)) were measured from RNA via real time PCR. Expression of tight junction protein 1 (TJP1) was measured in RNA isolated from ARPE-19s, to assess barrier stability. Results: The triple-culture promotes certain cell functionality through up-regulation of TJP1, increasing PEDF and decreasing VEGF expression highlighting its importance for the assessment of disease mechanisms distinct from a solo culture which would not allow the true effect of the native microenvironment to be elucidated. Conclusion: This model's novelty and reliability allows for the assessment of singular cellular function within the retinal microenvironment and overall assessment of retinal health, whilst eliminating the requirement of animal-based models. Journal Article Clinical & Experimental Ophthalmology 1442-6404 1442-9071 Retina; Retinal model; RPE biology; 31 12 2019 2019-12-31 10.1111/ceo.13578 COLLEGE NANME Sports Science COLLEGE CODE STSC Swansea University 2019-08-15T13:19:07.7882121 2019-07-01T08:18:05.6180713 Rachel Churm 0000-0001-9855-6282 1 Gareth J. Dunseath 2 Sarah Prior 0000-0001-8703-8092 3 Rebecca L. Thomas 4 Sanjiv Banerjee 5 David Owens 0000-0003-1002-1238 6 Becky Thomas 7 0050970-17072019111927.pdf 50970.pdf 2019-07-17T11:19:27.4930000 Output 724829 application/pdf Accepted Manuscript true 2020-06-29T00:00:00.0000000 true eng
title Development and characterization of an in vitro system of the human retina using cultured cell lines
spellingShingle Development and characterization of an in vitro system of the human retina using cultured cell lines
Rachel, Churm
Sarah, Prior
David, Owens
Becky, Thomas
title_short Development and characterization of an in vitro system of the human retina using cultured cell lines
title_full Development and characterization of an in vitro system of the human retina using cultured cell lines
title_fullStr Development and characterization of an in vitro system of the human retina using cultured cell lines
title_full_unstemmed Development and characterization of an in vitro system of the human retina using cultured cell lines
title_sort Development and characterization of an in vitro system of the human retina using cultured cell lines
author_id_str_mv c6cd8267ff0b13f2ea333bbfefdae144
cdda101035997acfaa6fdf17097f52b2
2fd4b7c3f82c6d3bd546eff61ff944e9
e83b45ec71428bd748ce201048f43d6a
author_id_fullname_str_mv c6cd8267ff0b13f2ea333bbfefdae144_***_Rachel, Churm
cdda101035997acfaa6fdf17097f52b2_***_Sarah, Prior
2fd4b7c3f82c6d3bd546eff61ff944e9_***_David, Owens
e83b45ec71428bd748ce201048f43d6a_***_Becky, Thomas
author Rachel, Churm
Sarah, Prior
David, Owens
Becky, Thomas
format Journal article
container_title Clinical & Experimental Ophthalmology
publishDate 2019
institution Swansea University
issn 1442-6404
1442-9071
doi_str_mv 10.1111/ceo.13578
document_store_str 1
active_str 0
description Background: Previously developed in vitro cultures of the human retina have been solo or dual cell cultures. We developed a triple-cell culture in vitro model utilizing a membrane system to produce a better representation of a functional and morphological human retina. Methods: Retinal microvascular endothelial cells (HRMVEC/ACBRI181, Cell systems), retinal pigment epithelium cells (RPE/ARPE-19, ATCC) and Müller glial cells (MIO-M1, UCL) were grown in a triple-culture. Our optimized triple-culture media contained a mix of specific endothelial medium and high glucose Dulbecco's Modified Eagle's medium (DMEM), where all three layers were viable for up to 5 days. Co-culture effect on morphological changes (cell staining) and gene expression of functional genes (pigment epithelial derived factor (PEDF) and vascular endothelial growth factor (VEGF)) were measured from RNA via real time PCR. Expression of tight junction protein 1 (TJP1) was measured in RNA isolated from ARPE-19s, to assess barrier stability. Results: The triple-culture promotes certain cell functionality through up-regulation of TJP1, increasing PEDF and decreasing VEGF expression highlighting its importance for the assessment of disease mechanisms distinct from a solo culture which would not allow the true effect of the native microenvironment to be elucidated. Conclusion: This model's novelty and reliability allows for the assessment of singular cellular function within the retinal microenvironment and overall assessment of retinal health, whilst eliminating the requirement of animal-based models.
published_date 2019-12-31T04:18:35Z
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