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Microfluidic co-cultures of retinal pigment epithelial cells and vascular endothelial cells to investigate choroidal angiogenesis

Angiogenesis plays a critical role in many diseases, including macular degeneration. At present, the pathological mechanisms remain unclear while appropriate models dissecting regulation of angiogenic processes are lacking. We propose an in vitro angiogenesis process and test it by examining the co-...

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Autores principales: Chen, Li-Jiun, Ito, Shuntaro, Kai, Hiroyuki, Nagamine, Kuniaki, Nagai, Nobuhiro, Nishizawa, Matsuhiko, Abe, Toshiaki, Kaji, Hirokazu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5471206/
https://www.ncbi.nlm.nih.gov/pubmed/28615726
http://dx.doi.org/10.1038/s41598-017-03788-5
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author Chen, Li-Jiun
Ito, Shuntaro
Kai, Hiroyuki
Nagamine, Kuniaki
Nagai, Nobuhiro
Nishizawa, Matsuhiko
Abe, Toshiaki
Kaji, Hirokazu
author_facet Chen, Li-Jiun
Ito, Shuntaro
Kai, Hiroyuki
Nagamine, Kuniaki
Nagai, Nobuhiro
Nishizawa, Matsuhiko
Abe, Toshiaki
Kaji, Hirokazu
author_sort Chen, Li-Jiun
collection PubMed
description Angiogenesis plays a critical role in many diseases, including macular degeneration. At present, the pathological mechanisms remain unclear while appropriate models dissecting regulation of angiogenic processes are lacking. We propose an in vitro angiogenesis process and test it by examining the co-culture of human retinal pigmental epithelial cells (ARPE-19) and human umbilical vein endothelial cells (HUVEC) inside a microfluidic device. From characterisation of the APRE-19 monoculture, the tight junction protein (ZO-1) was found on the cells cultured in the microfluidic device but changes in the medium conditions did not affect the integrity of monolayers found in the permeability tests. Vascular endothelial growth factor (VEGF) secretion was elevated under low glucose and hypoxia conditions compared to the control. After confirming the angiogenic ability of HUVEC, the cell-cell interactions were analyzed under lowered glucose medium and chemical hypoxia by exposing ARPE-19 cells to cobalt (II) chloride (CoCl(2)). Heterotypic interactions between ARPE-19 and HUVEC were observed, but proliferation of HUVEC was hindered once the monolayer of ARPE-19 started breaking down. The above characterisations showed that alterations in glucose concentration and/or oxygen level as induced by chemical hypoxia causes elevations in VEGF produced in ARPE-19 which in turn affected directional growth of HUVEC.
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spelling pubmed-54712062017-06-19 Microfluidic co-cultures of retinal pigment epithelial cells and vascular endothelial cells to investigate choroidal angiogenesis Chen, Li-Jiun Ito, Shuntaro Kai, Hiroyuki Nagamine, Kuniaki Nagai, Nobuhiro Nishizawa, Matsuhiko Abe, Toshiaki Kaji, Hirokazu Sci Rep Article Angiogenesis plays a critical role in many diseases, including macular degeneration. At present, the pathological mechanisms remain unclear while appropriate models dissecting regulation of angiogenic processes are lacking. We propose an in vitro angiogenesis process and test it by examining the co-culture of human retinal pigmental epithelial cells (ARPE-19) and human umbilical vein endothelial cells (HUVEC) inside a microfluidic device. From characterisation of the APRE-19 monoculture, the tight junction protein (ZO-1) was found on the cells cultured in the microfluidic device but changes in the medium conditions did not affect the integrity of monolayers found in the permeability tests. Vascular endothelial growth factor (VEGF) secretion was elevated under low glucose and hypoxia conditions compared to the control. After confirming the angiogenic ability of HUVEC, the cell-cell interactions were analyzed under lowered glucose medium and chemical hypoxia by exposing ARPE-19 cells to cobalt (II) chloride (CoCl(2)). Heterotypic interactions between ARPE-19 and HUVEC were observed, but proliferation of HUVEC was hindered once the monolayer of ARPE-19 started breaking down. The above characterisations showed that alterations in glucose concentration and/or oxygen level as induced by chemical hypoxia causes elevations in VEGF produced in ARPE-19 which in turn affected directional growth of HUVEC. Nature Publishing Group UK 2017-06-14 /pmc/articles/PMC5471206/ /pubmed/28615726 http://dx.doi.org/10.1038/s41598-017-03788-5 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Chen, Li-Jiun
Ito, Shuntaro
Kai, Hiroyuki
Nagamine, Kuniaki
Nagai, Nobuhiro
Nishizawa, Matsuhiko
Abe, Toshiaki
Kaji, Hirokazu
Microfluidic co-cultures of retinal pigment epithelial cells and vascular endothelial cells to investigate choroidal angiogenesis
title Microfluidic co-cultures of retinal pigment epithelial cells and vascular endothelial cells to investigate choroidal angiogenesis
title_full Microfluidic co-cultures of retinal pigment epithelial cells and vascular endothelial cells to investigate choroidal angiogenesis
title_fullStr Microfluidic co-cultures of retinal pigment epithelial cells and vascular endothelial cells to investigate choroidal angiogenesis
title_full_unstemmed Microfluidic co-cultures of retinal pigment epithelial cells and vascular endothelial cells to investigate choroidal angiogenesis
title_short Microfluidic co-cultures of retinal pigment epithelial cells and vascular endothelial cells to investigate choroidal angiogenesis
title_sort microfluidic co-cultures of retinal pigment epithelial cells and vascular endothelial cells to investigate choroidal angiogenesis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5471206/
https://www.ncbi.nlm.nih.gov/pubmed/28615726
http://dx.doi.org/10.1038/s41598-017-03788-5
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