Laminin-511-E8 promotes efficient in vitro expansion of human limbal melanocytes

Abstract Limbal melanocytes, located in the basal epithelial layer of the corneoscleral limbus, represent essential components of the corneal epithelial stem cell niche, but, due to difficulties in their isolation and cultivation, their biological roles and potential for stem cell-based tissue engin...

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Autores principales:	Naresh Polisetti, Andreas Gießl, Shen Li, Lydia Sorokin, Friedrich E. Kruse, Ursula Schlötzer-Schrehardt
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/702f27b969ae4b88a95fb558404cf9ee
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spelling	oai:doaj.org-article:702f27b969ae4b88a95fb558404cf9ee2021-12-02T15:39:57ZLaminin-511-E8 promotes efficient in vitro expansion of human limbal melanocytes10.1038/s41598-020-68120-02045-2322https://doaj.org/article/702f27b969ae4b88a95fb558404cf9ee2020-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-68120-0https://doaj.org/toc/2045-2322Abstract Limbal melanocytes, located in the basal epithelial layer of the corneoscleral limbus, represent essential components of the corneal epithelial stem cell niche, but, due to difficulties in their isolation and cultivation, their biological roles and potential for stem cell-based tissue engineering approaches have not been comprehensively studied. Here, we established a protocol for the efficient isolation and cultivation of pure populations of human limbal melanocytes, which could be expanded at high yield by using recombinant laminin (LN)-511-E8 as culture substrate. Co-cultivation of limbal melanocytes with limbal epithelial stem/progenitor cells on fibrin hydrogels pre-incubated with LN-511-E8 resulted in multilayered stratified epithelial constructs within ten days. By reproducing physiological cell–cell and cell–matrix interactions of the native niche environment, these biomimetic co-culture systems provide a promising experimental model for investigating the functional roles of melanocytes in the limbal stem cell niche and their suitability for developing advanced epithelial grafts for ocular surface surface reconstruction.Naresh PolisettiAndreas GießlShen LiLydia SorokinFriedrich E. KruseUrsula Schlötzer-SchrehardtNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-14 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Naresh Polisetti Andreas Gießl Shen Li Lydia Sorokin Friedrich E. Kruse Ursula Schlötzer-Schrehardt Laminin-511-E8 promotes efficient in vitro expansion of human limbal melanocytes
description	Abstract Limbal melanocytes, located in the basal epithelial layer of the corneoscleral limbus, represent essential components of the corneal epithelial stem cell niche, but, due to difficulties in their isolation and cultivation, their biological roles and potential for stem cell-based tissue engineering approaches have not been comprehensively studied. Here, we established a protocol for the efficient isolation and cultivation of pure populations of human limbal melanocytes, which could be expanded at high yield by using recombinant laminin (LN)-511-E8 as culture substrate. Co-cultivation of limbal melanocytes with limbal epithelial stem/progenitor cells on fibrin hydrogels pre-incubated with LN-511-E8 resulted in multilayered stratified epithelial constructs within ten days. By reproducing physiological cell–cell and cell–matrix interactions of the native niche environment, these biomimetic co-culture systems provide a promising experimental model for investigating the functional roles of melanocytes in the limbal stem cell niche and their suitability for developing advanced epithelial grafts for ocular surface surface reconstruction.
format	article
author	Naresh Polisetti Andreas Gießl Shen Li Lydia Sorokin Friedrich E. Kruse Ursula Schlötzer-Schrehardt
author_facet	Naresh Polisetti Andreas Gießl Shen Li Lydia Sorokin Friedrich E. Kruse Ursula Schlötzer-Schrehardt
author_sort	Naresh Polisetti
title	Laminin-511-E8 promotes efficient in vitro expansion of human limbal melanocytes
title_short	Laminin-511-E8 promotes efficient in vitro expansion of human limbal melanocytes
title_full	Laminin-511-E8 promotes efficient in vitro expansion of human limbal melanocytes
title_fullStr	Laminin-511-E8 promotes efficient in vitro expansion of human limbal melanocytes
title_full_unstemmed	Laminin-511-E8 promotes efficient in vitro expansion of human limbal melanocytes
title_sort	laminin-511-e8 promotes efficient in vitro expansion of human limbal melanocytes
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/702f27b969ae4b88a95fb558404cf9ee
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_version_	1718385879615012864

Laminin-511-E8 promotes efficient in vitro expansion of human limbal melanocytes

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