Culture surface protein coatings affect the barrier properties and calcium signalling of hESC-RPE

Abstract Human pluripotent stem cell-derived retinal pigment epithelium (RPE) transplantation is currently under evaluation as treatment for macular degeneration. For therapeutic applications, cryostorage during cell production is typically needed with potential consequences to cell functionality. W...

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Autores principales:	Taina Viheriälä, Juhana Sorvari, Teemu O. Ihalainen, Anni Mörö, Pyry Grönroos, Sabrina Schlie-Wolter, Boris Chichkov, Heli Skottman, Soile Nymark, Tanja Ilmarinen
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d31163423f174aa6ae385f815f57c54c
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spelling	oai:doaj.org-article:d31163423f174aa6ae385f815f57c54c2021-12-02T14:01:36ZCulture surface protein coatings affect the barrier properties and calcium signalling of hESC-RPE10.1038/s41598-020-79638-82045-2322https://doaj.org/article/d31163423f174aa6ae385f815f57c54c2021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-79638-8https://doaj.org/toc/2045-2322Abstract Human pluripotent stem cell-derived retinal pigment epithelium (RPE) transplantation is currently under evaluation as treatment for macular degeneration. For therapeutic applications, cryostorage during cell production is typically needed with potential consequences to cell functionality. We have previously shown that the culture substrate affects human embryonic stem cell-derived RPE (hESC-RPE) properties in fresh cultures. Here, we aimed to further identify the role of RPE basement membrane proteins type IV collagen (Col-IV), laminin (LN), and nidogen-1 in the maturation and functionality of hESC-RPE after cryopreservation. In addition to cell attachment and morphology, transepithelial electrical resistance, expression of key RPE proteins, phagocytosis capacity and Ca2+ signalling were analysed. After cryostorage, attachment of hESC-RPE on culture surfaces coated with Col-IV alone was poor. Combining Col-IV and LN with or without nidogen-1 significantly improved cell attachment and barrier properties of the epithelium. Furthermore, functional homogeneity of the hESC-RPE monolayer was enhanced in the presence of nidogen-1. Our results suggest that the choice of coating proteins for the cell culture may have implications to the functional properties of these cells after cryostorage cell banking.Taina ViheriäläJuhana SorvariTeemu O. IhalainenAnni MöröPyry GrönroosSabrina Schlie-WolterBoris ChichkovHeli SkottmanSoile NymarkTanja IlmarinenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Taina Viheriälä Juhana Sorvari Teemu O. Ihalainen Anni Mörö Pyry Grönroos Sabrina Schlie-Wolter Boris Chichkov Heli Skottman Soile Nymark Tanja Ilmarinen Culture surface protein coatings affect the barrier properties and calcium signalling of hESC-RPE
description	Abstract Human pluripotent stem cell-derived retinal pigment epithelium (RPE) transplantation is currently under evaluation as treatment for macular degeneration. For therapeutic applications, cryostorage during cell production is typically needed with potential consequences to cell functionality. We have previously shown that the culture substrate affects human embryonic stem cell-derived RPE (hESC-RPE) properties in fresh cultures. Here, we aimed to further identify the role of RPE basement membrane proteins type IV collagen (Col-IV), laminin (LN), and nidogen-1 in the maturation and functionality of hESC-RPE after cryopreservation. In addition to cell attachment and morphology, transepithelial electrical resistance, expression of key RPE proteins, phagocytosis capacity and Ca2+ signalling were analysed. After cryostorage, attachment of hESC-RPE on culture surfaces coated with Col-IV alone was poor. Combining Col-IV and LN with or without nidogen-1 significantly improved cell attachment and barrier properties of the epithelium. Furthermore, functional homogeneity of the hESC-RPE monolayer was enhanced in the presence of nidogen-1. Our results suggest that the choice of coating proteins for the cell culture may have implications to the functional properties of these cells after cryostorage cell banking.
format	article
author	Taina Viheriälä Juhana Sorvari Teemu O. Ihalainen Anni Mörö Pyry Grönroos Sabrina Schlie-Wolter Boris Chichkov Heli Skottman Soile Nymark Tanja Ilmarinen
author_facet	Taina Viheriälä Juhana Sorvari Teemu O. Ihalainen Anni Mörö Pyry Grönroos Sabrina Schlie-Wolter Boris Chichkov Heli Skottman Soile Nymark Tanja Ilmarinen
author_sort	Taina Viheriälä
title	Culture surface protein coatings affect the barrier properties and calcium signalling of hESC-RPE
title_short	Culture surface protein coatings affect the barrier properties and calcium signalling of hESC-RPE
title_full	Culture surface protein coatings affect the barrier properties and calcium signalling of hESC-RPE
title_fullStr	Culture surface protein coatings affect the barrier properties and calcium signalling of hESC-RPE
title_full_unstemmed	Culture surface protein coatings affect the barrier properties and calcium signalling of hESC-RPE
title_sort	culture surface protein coatings affect the barrier properties and calcium signalling of hesc-rpe
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/d31163423f174aa6ae385f815f57c54c
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Culture surface protein coatings affect the barrier properties and calcium signalling of hESC-RPE

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