Phenotype changes of oral epithelial stem cells after in vitro culture

Detalhes bibliográficos
Autor(a) principal: DALTOE,Felipe Perozzo
Data de Publicação: 2020
Outros Autores: OLIVEIRA,Nélio Alessandro Jesus de, PERON,Cibele Nunes, SHARPE,Paul Thomas, MANTESSO,Andrea
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Oral Research
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1806-83242020000100223
Resumo: Abstract The aim of our study was to isolate populations of keratinocyte stem cells based on the expression of cell surface markers and to investigate whether the culture could affect their phenotype. keratinocytes from human oral mucosa were sorted based on the expression of the epithelial stem cell markers p75NTR and CD71. We also examined the co-expression of other epithelial stem markers such as integrins β1 and α6 and their stem cell-like proprieties in in vitro assays. Three passages after being sorted by MACS, more than 93% of the p75NTR+ve cells lost the expression of p75NTR, while 5.46% of the p75NTR-ve gained it. Within the small population of the p75NTR+ve cells, 88% co-expressed other epithelial stem cell markers such as integrins β1 and α6, while only 28% of p75NTR-ve cells co-expressed these markers. These results were confirmed by sorting cells by FACS. Additionally, when double staining was used for sorting cells, 99% of the p75NTR+veCD71-ve and 33% of the p75NTR-veCD71+ve cells expressed both integrins, but just one week after culture, only 1.74% of the p75NTR+veCD71-ve cells still expressed p75NTR and only 0.32% still expressed CD71. Similar results were obtained when co-culturing p75NTR+ve and p75NTR-ve populations before analysis. Our results suggest that phenotype changes may be part of an intrinsic cellular mechanism to conserve levels of protein expression as they may found in the human body. In addition, in vitro culture may not offer ideal conditions for epithelial stem cell maintenance due to phenotype changes under standard culture conditions.
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spelling Phenotype changes of oral epithelial stem cells after in vitro cultureStem CellsMouth MucosaAbstract The aim of our study was to isolate populations of keratinocyte stem cells based on the expression of cell surface markers and to investigate whether the culture could affect their phenotype. keratinocytes from human oral mucosa were sorted based on the expression of the epithelial stem cell markers p75NTR and CD71. We also examined the co-expression of other epithelial stem markers such as integrins β1 and α6 and their stem cell-like proprieties in in vitro assays. Three passages after being sorted by MACS, more than 93% of the p75NTR+ve cells lost the expression of p75NTR, while 5.46% of the p75NTR-ve gained it. Within the small population of the p75NTR+ve cells, 88% co-expressed other epithelial stem cell markers such as integrins β1 and α6, while only 28% of p75NTR-ve cells co-expressed these markers. These results were confirmed by sorting cells by FACS. Additionally, when double staining was used for sorting cells, 99% of the p75NTR+veCD71-ve and 33% of the p75NTR-veCD71+ve cells expressed both integrins, but just one week after culture, only 1.74% of the p75NTR+veCD71-ve cells still expressed p75NTR and only 0.32% still expressed CD71. Similar results were obtained when co-culturing p75NTR+ve and p75NTR-ve populations before analysis. Our results suggest that phenotype changes may be part of an intrinsic cellular mechanism to conserve levels of protein expression as they may found in the human body. In addition, in vitro culture may not offer ideal conditions for epithelial stem cell maintenance due to phenotype changes under standard culture conditions.Sociedade Brasileira de Pesquisa Odontológica - SBPqO2020-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1806-83242020000100223Brazilian Oral Research v.34 2020reponame:Brazilian Oral Researchinstname:Sociedade Brasileira de Pesquisa Odontológica (SBPqO)instacron:SBPQO10.1590/1807-3107bor-2020.vol34.0033info:eu-repo/semantics/openAccessDALTOE,Felipe PerozzoOLIVEIRA,Nélio Alessandro Jesus dePERON,Cibele NunesSHARPE,Paul ThomasMANTESSO,Andreaeng2020-04-02T00:00:00Zoai:scielo:S1806-83242020000100223Revistahttps://www.scielo.br/j/bor/https://old.scielo.br/oai/scielo-oai.phppob@edu.usp.br||bor@sbpqo.org.br1807-31071806-8324opendoar:2020-04-02T00:00Brazilian Oral Research - Sociedade Brasileira de Pesquisa Odontológica (SBPqO)false
dc.title.none.fl_str_mv Phenotype changes of oral epithelial stem cells after in vitro culture
title Phenotype changes of oral epithelial stem cells after in vitro culture
spellingShingle Phenotype changes of oral epithelial stem cells after in vitro culture
DALTOE,Felipe Perozzo
Stem Cells
Mouth Mucosa
title_short Phenotype changes of oral epithelial stem cells after in vitro culture
title_full Phenotype changes of oral epithelial stem cells after in vitro culture
title_fullStr Phenotype changes of oral epithelial stem cells after in vitro culture
title_full_unstemmed Phenotype changes of oral epithelial stem cells after in vitro culture
title_sort Phenotype changes of oral epithelial stem cells after in vitro culture
author DALTOE,Felipe Perozzo
author_facet DALTOE,Felipe Perozzo
OLIVEIRA,Nélio Alessandro Jesus de
PERON,Cibele Nunes
SHARPE,Paul Thomas
MANTESSO,Andrea
author_role author
author2 OLIVEIRA,Nélio Alessandro Jesus de
PERON,Cibele Nunes
SHARPE,Paul Thomas
MANTESSO,Andrea
author2_role author
author
author
author
dc.contributor.author.fl_str_mv DALTOE,Felipe Perozzo
OLIVEIRA,Nélio Alessandro Jesus de
PERON,Cibele Nunes
SHARPE,Paul Thomas
MANTESSO,Andrea
dc.subject.por.fl_str_mv Stem Cells
Mouth Mucosa
topic Stem Cells
Mouth Mucosa
description Abstract The aim of our study was to isolate populations of keratinocyte stem cells based on the expression of cell surface markers and to investigate whether the culture could affect their phenotype. keratinocytes from human oral mucosa were sorted based on the expression of the epithelial stem cell markers p75NTR and CD71. We also examined the co-expression of other epithelial stem markers such as integrins β1 and α6 and their stem cell-like proprieties in in vitro assays. Three passages after being sorted by MACS, more than 93% of the p75NTR+ve cells lost the expression of p75NTR, while 5.46% of the p75NTR-ve gained it. Within the small population of the p75NTR+ve cells, 88% co-expressed other epithelial stem cell markers such as integrins β1 and α6, while only 28% of p75NTR-ve cells co-expressed these markers. These results were confirmed by sorting cells by FACS. Additionally, when double staining was used for sorting cells, 99% of the p75NTR+veCD71-ve and 33% of the p75NTR-veCD71+ve cells expressed both integrins, but just one week after culture, only 1.74% of the p75NTR+veCD71-ve cells still expressed p75NTR and only 0.32% still expressed CD71. Similar results were obtained when co-culturing p75NTR+ve and p75NTR-ve populations before analysis. Our results suggest that phenotype changes may be part of an intrinsic cellular mechanism to conserve levels of protein expression as they may found in the human body. In addition, in vitro culture may not offer ideal conditions for epithelial stem cell maintenance due to phenotype changes under standard culture conditions.
publishDate 2020
dc.date.none.fl_str_mv 2020-01-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1806-83242020000100223
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1806-83242020000100223
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/1807-3107bor-2020.vol34.0033
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Sociedade Brasileira de Pesquisa Odontológica - SBPqO
publisher.none.fl_str_mv Sociedade Brasileira de Pesquisa Odontológica - SBPqO
dc.source.none.fl_str_mv Brazilian Oral Research v.34 2020
reponame:Brazilian Oral Research
instname:Sociedade Brasileira de Pesquisa Odontológica (SBPqO)
instacron:SBPQO
instname_str Sociedade Brasileira de Pesquisa Odontológica (SBPqO)
instacron_str SBPQO
institution SBPQO
reponame_str Brazilian Oral Research
collection Brazilian Oral Research
repository.name.fl_str_mv Brazilian Oral Research - Sociedade Brasileira de Pesquisa Odontológica (SBPqO)
repository.mail.fl_str_mv pob@edu.usp.br||bor@sbpqo.org.br
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