Isolation and enrichment of melanocytes from human corneal limbus using CD117 (c-Kit) as selection marker

Limbal melanocytes (LM) are located in the basal epithelial layer of the corneoscleral limbus and interact with adjacent limbal epithelial progenitor cells. The exploration of their biological role in the maintenance of the limbal stem cell niche has been limited by the difficulty of LM isolation an...

Descripción completa

Detalles Bibliográficos
Autores principales: Polisetti, Naresh, Schlötzer-Schrehardt, Ursula, Reinhard, Thomas, Schlunck, Günther
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7567782/
https://www.ncbi.nlm.nih.gov/pubmed/33067486
http://dx.doi.org/10.1038/s41598-020-74869-1
_version_ 1783596394374758400
author Polisetti, Naresh
Schlötzer-Schrehardt, Ursula
Reinhard, Thomas
Schlunck, Günther
author_facet Polisetti, Naresh
Schlötzer-Schrehardt, Ursula
Reinhard, Thomas
Schlunck, Günther
author_sort Polisetti, Naresh
collection PubMed
description Limbal melanocytes (LM) are located in the basal epithelial layer of the corneoscleral limbus and interact with adjacent limbal epithelial progenitor cells. The exploration of their biological role in the maintenance of the limbal stem cell niche has been limited by the difficulty of LM isolation and cultivation. Here, we report on a facile protocol for the efficient isolation and enrichment of pure populations of human LMs by fluorescence-activated cell sorting (FACS) using antibodies raised against the cell surface marker CD117 (c-Kit). The enriched LMs retain self-renewal capacity and sustained melanin production, and are suitable to study the potential of LMs in stem cell-based corneal tissue engineering.
format Online
Article
Text
id pubmed-7567782
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-75677822020-10-19 Isolation and enrichment of melanocytes from human corneal limbus using CD117 (c-Kit) as selection marker Polisetti, Naresh Schlötzer-Schrehardt, Ursula Reinhard, Thomas Schlunck, Günther Sci Rep Article Limbal melanocytes (LM) are located in the basal epithelial layer of the corneoscleral limbus and interact with adjacent limbal epithelial progenitor cells. The exploration of their biological role in the maintenance of the limbal stem cell niche has been limited by the difficulty of LM isolation and cultivation. Here, we report on a facile protocol for the efficient isolation and enrichment of pure populations of human LMs by fluorescence-activated cell sorting (FACS) using antibodies raised against the cell surface marker CD117 (c-Kit). The enriched LMs retain self-renewal capacity and sustained melanin production, and are suitable to study the potential of LMs in stem cell-based corneal tissue engineering. Nature Publishing Group UK 2020-10-16 /pmc/articles/PMC7567782/ /pubmed/33067486 http://dx.doi.org/10.1038/s41598-020-74869-1 Text en © The Author(s) 2020 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Polisetti, Naresh
Schlötzer-Schrehardt, Ursula
Reinhard, Thomas
Schlunck, Günther
Isolation and enrichment of melanocytes from human corneal limbus using CD117 (c-Kit) as selection marker
title Isolation and enrichment of melanocytes from human corneal limbus using CD117 (c-Kit) as selection marker
title_full Isolation and enrichment of melanocytes from human corneal limbus using CD117 (c-Kit) as selection marker
title_fullStr Isolation and enrichment of melanocytes from human corneal limbus using CD117 (c-Kit) as selection marker
title_full_unstemmed Isolation and enrichment of melanocytes from human corneal limbus using CD117 (c-Kit) as selection marker
title_short Isolation and enrichment of melanocytes from human corneal limbus using CD117 (c-Kit) as selection marker
title_sort isolation and enrichment of melanocytes from human corneal limbus using cd117 (c-kit) as selection marker
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7567782/
https://www.ncbi.nlm.nih.gov/pubmed/33067486
http://dx.doi.org/10.1038/s41598-020-74869-1
work_keys_str_mv AT polisettinaresh isolationandenrichmentofmelanocytesfromhumancorneallimbususingcd117ckitasselectionmarker
AT schlotzerschrehardtursula isolationandenrichmentofmelanocytesfromhumancorneallimbususingcd117ckitasselectionmarker
AT reinhardthomas isolationandenrichmentofmelanocytesfromhumancorneallimbususingcd117ckitasselectionmarker
AT schlunckgunther isolationandenrichmentofmelanocytesfromhumancorneallimbususingcd117ckitasselectionmarker

Ejemplares similares