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

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...

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Autores principales: Polisetti, Naresh, Sorokin, Lydia, Okumura, Naoki, Koizumi, Noriko, Kinoshita, Shigeru, Kruse, Friedrich E., Schlötzer-Schrehardt, Ursula
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5506065/
https://www.ncbi.nlm.nih.gov/pubmed/28698551
http://dx.doi.org/10.1038/s41598-017-04916-x
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author Polisetti, Naresh
Sorokin, Lydia
Okumura, Naoki
Koizumi, Noriko
Kinoshita, Shigeru
Kruse, Friedrich E.
Schlötzer-Schrehardt, Ursula
author_facet Polisetti, Naresh
Sorokin, Lydia
Okumura, Naoki
Koizumi, Noriko
Kinoshita, Shigeru
Kruse, Friedrich E.
Schlötzer-Schrehardt, Ursula
author_sort Polisetti, Naresh
collection PubMed
description 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.
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spelling pubmed-55060652017-07-13 Laminin-511 and -521-based matrices for efficient ex vivo-expansion of human limbal epithelial progenitor cells Polisetti, Naresh Sorokin, Lydia Okumura, Naoki Koizumi, Noriko Kinoshita, Shigeru Kruse, Friedrich E. Schlötzer-Schrehardt, Ursula Sci Rep Article 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. Nature Publishing Group UK 2017-07-11 /pmc/articles/PMC5506065/ /pubmed/28698551 http://dx.doi.org/10.1038/s41598-017-04916-x 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
Polisetti, Naresh
Sorokin, Lydia
Okumura, Naoki
Koizumi, Noriko
Kinoshita, Shigeru
Kruse, Friedrich E.
Schlötzer-Schrehardt, Ursula
Laminin-511 and -521-based matrices for efficient ex vivo-expansion of human limbal epithelial progenitor cells
title 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_short 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
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5506065/
https://www.ncbi.nlm.nih.gov/pubmed/28698551
http://dx.doi.org/10.1038/s41598-017-04916-x
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