Laminin-511 and -521-based matrices for efficient ex vivo-expansion of human limbal epithelial progenitor cells

Abstract Optimization of culture conditions for human limbal epithelial stem/progenitor cells (LEPC) that incorporate the in vivo cell-matrix interactions are essential to enhance LEPC ex vivo-expansion and transplantation efficiency. Here, we investigate the efficacy of laminin (LN) isoforms prefer...

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Autores principales:	Naresh Polisetti, Lydia Sorokin, Naoki Okumura, Noriko Koizumi, Shigeru Kinoshita, Friedrich E. Kruse, Ursula Schlötzer-Schrehardt
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1507635635b5490a96657eab0f81eef8
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spelling	oai:doaj.org-article:1507635635b5490a96657eab0f81eef82021-12-02T16:07:59ZLaminin-511 and -521-based matrices for efficient ex vivo-expansion of human limbal epithelial progenitor cells10.1038/s41598-017-04916-x2045-2322https://doaj.org/article/1507635635b5490a96657eab0f81eef82017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-04916-xhttps://doaj.org/toc/2045-2322Abstract Optimization of culture conditions for human limbal epithelial stem/progenitor cells (LEPC) that incorporate the in vivo cell-matrix interactions are essential to enhance LEPC ex vivo-expansion and transplantation efficiency. Here, we investigate the efficacy of laminin (LN) isoforms preferentially expressed in the limbal niche as culture matrices for epithelial tissue engineering. Analyses of expression patterns of LN chains in the human limbal niche provided evidence for enrichment of LN-α2, -α3, -α5, -β1, -β2, -β3, -γ1, -γ2 and -γ3 chains in the limbal basement membrane, with LN-α5 representing a signature component specifically produced by epithelial progenitor cells. Recombinant human LN-521 and LN-511 significantly enhanced in vitro LEPC adhesion, migration and proliferation compared to other isoforms, and maintained phenotype stability. The bioactive LN-511-E8 fragment carrying only C-terminal domains showed similar efficacy as full-length LN-511. Functional blocking of α3β1 and α6β1 integrins suppressed adhesion of LEPC to LN-511/521-coated surfaces. Cultivation of LEPC on fibrin-based hydrogels incorporating LN-511-E8 resulted in firm integrin-mediated adhesion to the scaffold and well-stratified epithelial constructs, with maintenance of a progenitor cell phenotype in their (supra)basal layers. Thus, the incorporation of chemically defined LN-511-E8 into biosynthetic scaffolds represents a promising approach for xeno-free corneal epithelial tissue engineering for ocular surface reconstruction.Naresh PolisettiLydia SorokinNaoki OkumuraNoriko KoizumiShigeru KinoshitaFriedrich E. KruseUrsula Schlötzer-SchrehardtNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-15 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Naresh Polisetti Lydia Sorokin Naoki Okumura Noriko Koizumi Shigeru Kinoshita Friedrich E. Kruse Ursula Schlötzer-Schrehardt Laminin-511 and -521-based matrices for efficient ex vivo-expansion of human limbal epithelial progenitor cells
description	Abstract Optimization of culture conditions for human limbal epithelial stem/progenitor cells (LEPC) that incorporate the in vivo cell-matrix interactions are essential to enhance LEPC ex vivo-expansion and transplantation efficiency. Here, we investigate the efficacy of laminin (LN) isoforms preferentially expressed in the limbal niche as culture matrices for epithelial tissue engineering. Analyses of expression patterns of LN chains in the human limbal niche provided evidence for enrichment of LN-α2, -α3, -α5, -β1, -β2, -β3, -γ1, -γ2 and -γ3 chains in the limbal basement membrane, with LN-α5 representing a signature component specifically produced by epithelial progenitor cells. Recombinant human LN-521 and LN-511 significantly enhanced in vitro LEPC adhesion, migration and proliferation compared to other isoforms, and maintained phenotype stability. The bioactive LN-511-E8 fragment carrying only C-terminal domains showed similar efficacy as full-length LN-511. Functional blocking of α3β1 and α6β1 integrins suppressed adhesion of LEPC to LN-511/521-coated surfaces. Cultivation of LEPC on fibrin-based hydrogels incorporating LN-511-E8 resulted in firm integrin-mediated adhesion to the scaffold and well-stratified epithelial constructs, with maintenance of a progenitor cell phenotype in their (supra)basal layers. Thus, the incorporation of chemically defined LN-511-E8 into biosynthetic scaffolds represents a promising approach for xeno-free corneal epithelial tissue engineering for ocular surface reconstruction.
format	article
author	Naresh Polisetti Lydia Sorokin Naoki Okumura Noriko Koizumi Shigeru Kinoshita Friedrich E. Kruse Ursula Schlötzer-Schrehardt
author_facet	Naresh Polisetti Lydia Sorokin Naoki Okumura Noriko Koizumi Shigeru Kinoshita Friedrich E. Kruse Ursula Schlötzer-Schrehardt
author_sort	Naresh Polisetti
title	Laminin-511 and -521-based matrices for efficient ex vivo-expansion of human limbal epithelial progenitor cells
title_short	Laminin-511 and -521-based matrices for efficient ex vivo-expansion of human limbal epithelial progenitor cells
title_full	Laminin-511 and -521-based matrices for efficient ex vivo-expansion of human limbal epithelial progenitor cells
title_fullStr	Laminin-511 and -521-based matrices for efficient ex vivo-expansion of human limbal epithelial progenitor cells
title_full_unstemmed	Laminin-511 and -521-based matrices for efficient ex vivo-expansion of human limbal epithelial progenitor cells
title_sort	laminin-511 and -521-based matrices for efficient ex vivo-expansion of human limbal epithelial progenitor cells
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/1507635635b5490a96657eab0f81eef8
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_version_	1718384639692767232

Laminin-511 and -521-based matrices for efficient ex vivo-expansion of human limbal epithelial progenitor cells

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