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P-Cadherin Is Expressed by Epithelial Progenitor Cells and Melanocytes in the Human Corneal Limbus

Interactions between limbal epithelial progenitor cells (LEPC) and surrounding niche cells, which include limbal mesenchymal stromal cells (LMSC) and melanocytes (LM), are essential for the maintenance of the limbal stem cell niche required for a transparent corneal surface. P-cadherin (P-cad) is a...

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Autores principales: Polisetti, Naresh, Sharaf, Lyne, Martin, Gottfried, Schlunck, Günther, Reinhard, Thomas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9221557/
https://www.ncbi.nlm.nih.gov/pubmed/35741104
http://dx.doi.org/10.3390/cells11121975
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author Polisetti, Naresh
Sharaf, Lyne
Martin, Gottfried
Schlunck, Günther
Reinhard, Thomas
author_facet Polisetti, Naresh
Sharaf, Lyne
Martin, Gottfried
Schlunck, Günther
Reinhard, Thomas
author_sort Polisetti, Naresh
collection PubMed
description Interactions between limbal epithelial progenitor cells (LEPC) and surrounding niche cells, which include limbal mesenchymal stromal cells (LMSC) and melanocytes (LM), are essential for the maintenance of the limbal stem cell niche required for a transparent corneal surface. P-cadherin (P-cad) is a critical stem cell niche adhesion molecule at various epithelial stem cell niches; however, conflicting observations were reported on the presence of P-cad in the limbal region. To explore this issue, we assessed the location and phenotype of P-cad(+) cells by confocal microscopy of human corneoscleral tissue. In subsequent fluorescence-activated cell sorting (FACS) experiments, we used antibodies against P-cad along with CD90 and CD117 for the enrichment of LEPC, LMSC and LM, respectively. The sorted cells were characterized by immunophenotyping and the repopulation of decellularized limbal scaffolds was evaluated. Our findings demonstrate that P-cad is expressed by epithelial progenitor cells as well as melanocytes in the human limbal epithelial stem cell niche. The modified flow sorting addressing P-cad as well as CD90 and CD117 yielded enriched LEPC (CD90(−)CD117(−)P-cad(+)) and pure populations of LMSC (CD90(+)CD117(−)P-cad(−)) and LM (CD90(−)CD117(+)P-cad(+)). The enriched LEPC showed the expression of epithelial progenitor markers and better colony-forming ability than their P-cad(−) counterparts. The cultured LEPC and LM exhibited P-cad expression at intercellular junctions and successfully repopulated decellularized limbal scaffolds. These data suggest that P-cad is a critical cell–cell adhesion molecule, connecting LEPC and LM, which may play an important role in the long-term maintenance of LEPC at the limbal stem cell niche; moreover, these findings led to further improvement of cell enrichment protocols to enhance the yield of LEPC.
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spelling pubmed-92215572022-06-24 P-Cadherin Is Expressed by Epithelial Progenitor Cells and Melanocytes in the Human Corneal Limbus Polisetti, Naresh Sharaf, Lyne Martin, Gottfried Schlunck, Günther Reinhard, Thomas Cells Article Interactions between limbal epithelial progenitor cells (LEPC) and surrounding niche cells, which include limbal mesenchymal stromal cells (LMSC) and melanocytes (LM), are essential for the maintenance of the limbal stem cell niche required for a transparent corneal surface. P-cadherin (P-cad) is a critical stem cell niche adhesion molecule at various epithelial stem cell niches; however, conflicting observations were reported on the presence of P-cad in the limbal region. To explore this issue, we assessed the location and phenotype of P-cad(+) cells by confocal microscopy of human corneoscleral tissue. In subsequent fluorescence-activated cell sorting (FACS) experiments, we used antibodies against P-cad along with CD90 and CD117 for the enrichment of LEPC, LMSC and LM, respectively. The sorted cells were characterized by immunophenotyping and the repopulation of decellularized limbal scaffolds was evaluated. Our findings demonstrate that P-cad is expressed by epithelial progenitor cells as well as melanocytes in the human limbal epithelial stem cell niche. The modified flow sorting addressing P-cad as well as CD90 and CD117 yielded enriched LEPC (CD90(−)CD117(−)P-cad(+)) and pure populations of LMSC (CD90(+)CD117(−)P-cad(−)) and LM (CD90(−)CD117(+)P-cad(+)). The enriched LEPC showed the expression of epithelial progenitor markers and better colony-forming ability than their P-cad(−) counterparts. The cultured LEPC and LM exhibited P-cad expression at intercellular junctions and successfully repopulated decellularized limbal scaffolds. These data suggest that P-cad is a critical cell–cell adhesion molecule, connecting LEPC and LM, which may play an important role in the long-term maintenance of LEPC at the limbal stem cell niche; moreover, these findings led to further improvement of cell enrichment protocols to enhance the yield of LEPC. MDPI 2022-06-20 /pmc/articles/PMC9221557/ /pubmed/35741104 http://dx.doi.org/10.3390/cells11121975 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Polisetti, Naresh
Sharaf, Lyne
Martin, Gottfried
Schlunck, Günther
Reinhard, Thomas
P-Cadherin Is Expressed by Epithelial Progenitor Cells and Melanocytes in the Human Corneal Limbus
title P-Cadherin Is Expressed by Epithelial Progenitor Cells and Melanocytes in the Human Corneal Limbus
title_full P-Cadherin Is Expressed by Epithelial Progenitor Cells and Melanocytes in the Human Corneal Limbus
title_fullStr P-Cadherin Is Expressed by Epithelial Progenitor Cells and Melanocytes in the Human Corneal Limbus
title_full_unstemmed P-Cadherin Is Expressed by Epithelial Progenitor Cells and Melanocytes in the Human Corneal Limbus
title_short P-Cadherin Is Expressed by Epithelial Progenitor Cells and Melanocytes in the Human Corneal Limbus
title_sort p-cadherin is expressed by epithelial progenitor cells and melanocytes in the human corneal limbus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9221557/
https://www.ncbi.nlm.nih.gov/pubmed/35741104
http://dx.doi.org/10.3390/cells11121975
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