In vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers

Abstract Corneal endothelial cells (CECs) are very important for the maintenance of corneal transparency. However, in vitro, CECs display limited proliferation and loss of phenotype via endothelial to mesenchymal transformation (EMT) and cellular senescence. In this study, we demonstrate that contin...

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Autores principales:	Shanyi Li, Yuting Han, Hao Lei, Yingxin Zeng, Zekai Cui, Qiaolang Zeng, Deliang Zhu, Ruiling Lian, Jun Zhang, Zhe Chen, Jiansu Chen
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8a8251148c264a3aaf2769a2d8f6941c
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spelling	oai:doaj.org-article:8a8251148c264a3aaf2769a2d8f6941c2021-12-02T11:52:42ZIn vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers10.1038/s41598-017-00914-12045-2322https://doaj.org/article/8a8251148c264a3aaf2769a2d8f6941c2017-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00914-1https://doaj.org/toc/2045-2322Abstract Corneal endothelial cells (CECs) are very important for the maintenance of corneal transparency. However, in vitro, CECs display limited proliferation and loss of phenotype via endothelial to mesenchymal transformation (EMT) and cellular senescence. In this study, we demonstrate that continuous supplementary nutrition using a perfusion culture bioreactor and three-dimensional (3D) spheroid culture can be used to improve CEC expansion in culture and to construct a tissue-engineered CEC layer. Compared with static culture, perfusion-derived CECs exhibited an increased proliferative ability as well as formed close cell-cell contact junctions and numerous surface microvilli. We also demonstrated that the CEC spheroid culture significantly down-regulated gene expression of the proliferation marker Ki67 and EMT-related markers Vimentin and α-SMA, whereas the gene expression level of the CEC marker ATP1A1 was significantly up-regulated. Furthermore, use of the perfusion system in conjunction with a spheroid culture on decellularized corneal scaffolds and collagen sheets promoted the generation of CEC monolayers as well as neo-synthesized ECM formation. This study also confirmed that a CEC spheroid culture on a curved collagen sheet with controlled physiological intraocular pressure could generate a CEC monolayer. Thus, our results show that the use of a perfusion system and 3D spheroid culture can promote CEC expansion and the construction of tissue-engineered corneal endothelial layers in vitro.Shanyi LiYuting HanHao LeiYingxin ZengZekai CuiQiaolang ZengDeliang ZhuRuiling LianJun ZhangZhe ChenJiansu ChenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-17 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Shanyi Li Yuting Han Hao Lei Yingxin Zeng Zekai Cui Qiaolang Zeng Deliang Zhu Ruiling Lian Jun Zhang Zhe Chen Jiansu Chen In vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers
description	Abstract Corneal endothelial cells (CECs) are very important for the maintenance of corneal transparency. However, in vitro, CECs display limited proliferation and loss of phenotype via endothelial to mesenchymal transformation (EMT) and cellular senescence. In this study, we demonstrate that continuous supplementary nutrition using a perfusion culture bioreactor and three-dimensional (3D) spheroid culture can be used to improve CEC expansion in culture and to construct a tissue-engineered CEC layer. Compared with static culture, perfusion-derived CECs exhibited an increased proliferative ability as well as formed close cell-cell contact junctions and numerous surface microvilli. We also demonstrated that the CEC spheroid culture significantly down-regulated gene expression of the proliferation marker Ki67 and EMT-related markers Vimentin and α-SMA, whereas the gene expression level of the CEC marker ATP1A1 was significantly up-regulated. Furthermore, use of the perfusion system in conjunction with a spheroid culture on decellularized corneal scaffolds and collagen sheets promoted the generation of CEC monolayers as well as neo-synthesized ECM formation. This study also confirmed that a CEC spheroid culture on a curved collagen sheet with controlled physiological intraocular pressure could generate a CEC monolayer. Thus, our results show that the use of a perfusion system and 3D spheroid culture can promote CEC expansion and the construction of tissue-engineered corneal endothelial layers in vitro.
format	article
author	Shanyi Li Yuting Han Hao Lei Yingxin Zeng Zekai Cui Qiaolang Zeng Deliang Zhu Ruiling Lian Jun Zhang Zhe Chen Jiansu Chen
author_facet	Shanyi Li Yuting Han Hao Lei Yingxin Zeng Zekai Cui Qiaolang Zeng Deliang Zhu Ruiling Lian Jun Zhang Zhe Chen Jiansu Chen
author_sort	Shanyi Li
title	In vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers
title_short	In vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers
title_full	In vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers
title_fullStr	In vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers
title_full_unstemmed	In vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers
title_sort	in vitro biomimetic platforms featuring a perfusion system and 3d spheroid culture promote the construction of tissue-engineered corneal endothelial layers
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/8a8251148c264a3aaf2769a2d8f6941c
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In vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers

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