Bioengineered Bruch's-like extracellular matrix promotes retinal pigment epithelial differentiation

In the eye, the retinal pigment epithelium (RPE) adheres to a complex protein matrix known as Bruch's membrane (BrM). The aim of this study was to provide enriched conditions for RPE cell culture through the production of a BrM-like matrix. Our hypothesis was that a human RPE cell line would de...

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Autores: McLenachan, Samuel, Hao, Erwei, Zhang, Dan, Zhang, Ling, Edel, Michael John, Chen, Fred K.
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2017
País:España
Recursos:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositório:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:2445/116251
Acesso em linha:https://hdl.handle.net/2445/116251
Access Level:Acceso aberto
Palavra-chave:Oftalmologia
Retina
Ophthalmology
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spelling Bioengineered Bruch's-like extracellular matrix promotes retinal pigment epithelial differentiationMcLenachan, SamuelHao, ErweiZhang, DanZhang, LingEdel, Michael JohnChen, Fred K.OftalmologiaRetinaOphthalmologyRetinaIn the eye, the retinal pigment epithelium (RPE) adheres to a complex protein matrix known as Bruch's membrane (BrM). The aim of this study was to provide enriched conditions for RPE cell culture through the production of a BrM-like matrix. Our hypothesis was that a human RPE cell line would deposit an extracellular matrix (ECM) resembling BrM. The composition and structure of ECM deposited by ARPE19 cells (ARPE19-ECM) was characterized. To produce ARPE19-ECM, ARPE19 cells were cultured in the presence dextran sulphate. ARPE19-ECM was decellularized using deoxycholate and characterized by immunostaining and western blot analysis. Primary human RPE and induced pluripotent stem cells were seeded onto ARPE19-ECM or geltrex coated surfaces and examined by microscopy or RT-PCR. Culture of ARPE19 cells with dextran sulphate promoted nuclear localization of SOX2, formation of tight junctions and deposition of ECM. ARPE19 cells deposited ECM proteins found in the inner layers of BrM, including fibronectin, vitronectin, collagens IV and V as well as laminin-alpha-5, but not those found in the middle elastic layer (elastin) or the outer layers (collagen VI). ARPE19-ECM promoted pigmentation in human RPE and pluripotent stem cell cultures. Expression of RPE65 was significantly increased on ARPE19-ECM compared with geltrex in differentiating pluripotent stem cell cultures. ARPE19 cells deposit ECM with a composition and structure similar to BrM in the retina. Molecular cues present in ARPE19-ECM promote the acquisition and maintenance of the RPE phenotype. Together, these results demonstrate a simple method for generating a BrM-like surface for enriched RPE cell cultures.Elsevier2017201720172017info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion8 p.application/pdfhttps://hdl.handle.net/2445/116251Articles publicats en revistes (Biomedicina)reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésReproducció del document publicat a: https://doi.org/10.1016/j.bbrep.2017.03.008Biochemistry and Biophysics Reports, 2017, vol. 10, p. 178-185https://doi.org/10.1016/j.bbrep.2017.03.008cc-by-nc-nd (c) McLenachan⁠, Samuel et al., 2017http://creativecommons.org/licenses/by-nc-nd/3.0/esinfo:eu-repo/semantics/openAccessoai:recercat.cat:2445/1162512026-05-29T05:05:01Z
dc.title.none.fl_str_mv Bioengineered Bruch's-like extracellular matrix promotes retinal pigment epithelial differentiation
title Bioengineered Bruch's-like extracellular matrix promotes retinal pigment epithelial differentiation
spellingShingle Bioengineered Bruch's-like extracellular matrix promotes retinal pigment epithelial differentiation
McLenachan, Samuel
Oftalmologia
Retina
Ophthalmology
Retina
title_short Bioengineered Bruch's-like extracellular matrix promotes retinal pigment epithelial differentiation
title_full Bioengineered Bruch's-like extracellular matrix promotes retinal pigment epithelial differentiation
title_fullStr Bioengineered Bruch's-like extracellular matrix promotes retinal pigment epithelial differentiation
title_full_unstemmed Bioengineered Bruch's-like extracellular matrix promotes retinal pigment epithelial differentiation
title_sort Bioengineered Bruch's-like extracellular matrix promotes retinal pigment epithelial differentiation
dc.creator.none.fl_str_mv McLenachan, Samuel
Hao, Erwei
Zhang, Dan
Zhang, Ling
Edel, Michael John
Chen, Fred K.
author McLenachan, Samuel
author_facet McLenachan, Samuel
Hao, Erwei
Zhang, Dan
Zhang, Ling
Edel, Michael John
Chen, Fred K.
author_role author
author2 Hao, Erwei
Zhang, Dan
Zhang, Ling
Edel, Michael John
Chen, Fred K.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Oftalmologia
Retina
Ophthalmology
Retina
topic Oftalmologia
Retina
Ophthalmology
Retina
description In the eye, the retinal pigment epithelium (RPE) adheres to a complex protein matrix known as Bruch's membrane (BrM). The aim of this study was to provide enriched conditions for RPE cell culture through the production of a BrM-like matrix. Our hypothesis was that a human RPE cell line would deposit an extracellular matrix (ECM) resembling BrM. The composition and structure of ECM deposited by ARPE19 cells (ARPE19-ECM) was characterized. To produce ARPE19-ECM, ARPE19 cells were cultured in the presence dextran sulphate. ARPE19-ECM was decellularized using deoxycholate and characterized by immunostaining and western blot analysis. Primary human RPE and induced pluripotent stem cells were seeded onto ARPE19-ECM or geltrex coated surfaces and examined by microscopy or RT-PCR. Culture of ARPE19 cells with dextran sulphate promoted nuclear localization of SOX2, formation of tight junctions and deposition of ECM. ARPE19 cells deposited ECM proteins found in the inner layers of BrM, including fibronectin, vitronectin, collagens IV and V as well as laminin-alpha-5, but not those found in the middle elastic layer (elastin) or the outer layers (collagen VI). ARPE19-ECM promoted pigmentation in human RPE and pluripotent stem cell cultures. Expression of RPE65 was significantly increased on ARPE19-ECM compared with geltrex in differentiating pluripotent stem cell cultures. ARPE19 cells deposit ECM with a composition and structure similar to BrM in the retina. Molecular cues present in ARPE19-ECM promote the acquisition and maintenance of the RPE phenotype. Together, these results demonstrate a simple method for generating a BrM-like surface for enriched RPE cell cultures.
publishDate 2017
dc.date.none.fl_str_mv 2017
2017
2017
2017
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/116251
url https://hdl.handle.net/2445/116251
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.1016/j.bbrep.2017.03.008
Biochemistry and Biophysics Reports, 2017, vol. 10, p. 178-185
https://doi.org/10.1016/j.bbrep.2017.03.008
dc.rights.none.fl_str_mv cc-by-nc-nd (c) McLenachan⁠, Samuel et al., 2017
http://creativecommons.org/licenses/by-nc-nd/3.0/es
info:eu-repo/semantics/openAccess
rights_invalid_str_mv cc-by-nc-nd (c) McLenachan⁠, Samuel et al., 2017
http://creativecommons.org/licenses/by-nc-nd/3.0/es
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 8 p.
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv Articles publicats en revistes (Biomedicina)
reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.228081

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