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Development and characterization of an in vitro system of the human retina using cultured cell lines

Rachel Churm Orcid Logo, Gareth Dunseath Orcid Logo, Sarah Prior Orcid Logo, Frankie Thomas, Sanjiv Banerjee, David Owens Orcid Logo

Clinical & Experimental Ophthalmology, Volume: 47, Issue: 8, Pages: 1055 - 1062

Swansea University Authors: Rachel Churm Orcid Logo, Gareth Dunseath Orcid Logo, Sarah Prior Orcid Logo, Frankie Thomas, David Owens Orcid Logo

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DOI (Published version): 10.1111/ceo.13578

Abstract

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: Wiley 2019
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URI: https://cronfa.swan.ac.uk/Record/cronfa50970
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Abstract: 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.
Keywords: cell culture, human retina, in vitro model, triple culture
Issue: 8
Start Page: 1055
End Page: 1062