Isolation and long-term expansion of murine epidermal stem-like cells.

Epidermis is the most outer layer of the skin and a physical barrier protecting the internal tissues from mechanical and environmental insults. The basal keratinocytes, which, through proliferation and differentiation, supply diverse cell types for epidermal homeostasis and injury repair. Sustainabl...

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Autores principales:	Jingjing Wang, Maureen Mongan, Xiang Zhang, Ying Xia
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3256a9eb1682497793e0e830fc74c9bb
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spelling	oai:doaj.org-article:3256a9eb1682497793e0e830fc74c9bb2021-12-02T20:06:55ZIsolation and long-term expansion of murine epidermal stem-like cells.1932-620310.1371/journal.pone.0254731https://doaj.org/article/3256a9eb1682497793e0e830fc74c9bb2021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0254731https://doaj.org/toc/1932-6203Epidermis is the most outer layer of the skin and a physical barrier protecting the internal tissues from mechanical and environmental insults. The basal keratinocytes, which, through proliferation and differentiation, supply diverse cell types for epidermal homeostasis and injury repair. Sustainable culture of murine keratinocyte, however, is a major obstacle. Here we developed murine keratinocyte lines using low-Ca2+ (0.06 mM) keratinocyte serum-free medium (KSFM-Ca2+) without feeder cells. Cells derived in this condition could be subcultured for >70 passages. They displayed basal epithelial cell morphology and expressed keratin (Krt) 14, but lacked the epithelial-characteristic intercellular junctions. Moreover, these cells could be adapted to grow in the Defined-KSFM (DKSFM) media containing 0.15 mM Ca2+, and the adapted cells established tight- and adherens-junctions and exhibited increased Krt1/10 expression while retained subculture capacity. Global gene expression studies showed cells derived in KSFM-Ca2+ media had enriched stem/proliferation markers and cells adapted in DKSFM media had epithelial progenitor signatures. Correspondingly, KSFM-Ca2+-derived cells exhibited a remarkable capacity of clonal expansion, whereas DKSFM-adapted cells could differentiate to suprabasal epithelial cell types in 3-dimentional (3D) organoids. The generation of stem-like murine keratinocyte lines and the conversion of these cells to epithelial progenitors capable of terminal differentiation provide the critically needed resources for skin research.Jingjing WangMaureen MonganXiang ZhangYing XiaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 7, p e0254731 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jingjing Wang Maureen Mongan Xiang Zhang Ying Xia Isolation and long-term expansion of murine epidermal stem-like cells.
description	Epidermis is the most outer layer of the skin and a physical barrier protecting the internal tissues from mechanical and environmental insults. The basal keratinocytes, which, through proliferation and differentiation, supply diverse cell types for epidermal homeostasis and injury repair. Sustainable culture of murine keratinocyte, however, is a major obstacle. Here we developed murine keratinocyte lines using low-Ca2+ (0.06 mM) keratinocyte serum-free medium (KSFM-Ca2+) without feeder cells. Cells derived in this condition could be subcultured for >70 passages. They displayed basal epithelial cell morphology and expressed keratin (Krt) 14, but lacked the epithelial-characteristic intercellular junctions. Moreover, these cells could be adapted to grow in the Defined-KSFM (DKSFM) media containing 0.15 mM Ca2+, and the adapted cells established tight- and adherens-junctions and exhibited increased Krt1/10 expression while retained subculture capacity. Global gene expression studies showed cells derived in KSFM-Ca2+ media had enriched stem/proliferation markers and cells adapted in DKSFM media had epithelial progenitor signatures. Correspondingly, KSFM-Ca2+-derived cells exhibited a remarkable capacity of clonal expansion, whereas DKSFM-adapted cells could differentiate to suprabasal epithelial cell types in 3-dimentional (3D) organoids. The generation of stem-like murine keratinocyte lines and the conversion of these cells to epithelial progenitors capable of terminal differentiation provide the critically needed resources for skin research.
format	article
author	Jingjing Wang Maureen Mongan Xiang Zhang Ying Xia
author_facet	Jingjing Wang Maureen Mongan Xiang Zhang Ying Xia
author_sort	Jingjing Wang
title	Isolation and long-term expansion of murine epidermal stem-like cells.
title_short	Isolation and long-term expansion of murine epidermal stem-like cells.
title_full	Isolation and long-term expansion of murine epidermal stem-like cells.
title_fullStr	Isolation and long-term expansion of murine epidermal stem-like cells.
title_full_unstemmed	Isolation and long-term expansion of murine epidermal stem-like cells.
title_sort	isolation and long-term expansion of murine epidermal stem-like cells.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/3256a9eb1682497793e0e830fc74c9bb
work_keys_str_mv	AT jingjingwang isolationandlongtermexpansionofmurineepidermalstemlikecells AT maureenmongan isolationandlongtermexpansionofmurineepidermalstemlikecells AT xiangzhang isolationandlongtermexpansionofmurineepidermalstemlikecells AT yingxia isolationandlongtermexpansionofmurineepidermalstemlikecells
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Isolation and long-term expansion of murine epidermal stem-like cells.

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