Aldefluor protocol to sort keratinocytes stem cells from skin

Detalhes bibliográficos
Autor(a) principal: Noronha,Samuel Marcos Ribeiro
Data de Publicação: 2017
Outros Autores: Gragnani,Alfredo, Pereira,Thiago Antônio Calado, Correa,Silvana Aparecida Alves, Bonucci,Jessica, Ferreira,Lydia Masako
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Acta Cirúrgica Brasileira (Online)
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0102-86502017001100984
Resumo: Abstract Purpose: To investigate the use Aldefluor® and N, N - Dimethylaminobenzaldehyde (DEAB) to design a protocol to sort keratinocyte stem cells from cultured keratinocytes from burned patients. Methods: Activated Aldefluor® aliquots were prepared and maintained at temperature between 2 to 8°C, or stored at -20°C. Next, the cells were collected following the standard protocol of sample preparation. Results: Best results were obtained with Aldefluor® 1.5µl and DEAB 15 µl for 1 x 106 cells, incubated at 37°C for 15 minutes. Flow cytometer range for keratinocyte stem cells separation was evaluated. There were 14.8% of stem cells separated in one sample of keratinocyte culture used to pattern the protocol. After being defined the ideal concentration, the same test pattern was performed in other keratinocyte samples. We observed a final mean of 10.8%. Conclusion: Aldefluor® has been shown as a favorable marking of epidermal keratinocyte stem cells for subsequent separation on a flow cytometer, with detection of 10.8% of epidermal keratinocyte stem cells, in this protocol.
id SBDPC-1_431a63ea14660f4eb455f6080e4de856
oai_identifier_str oai:scielo:S0102-86502017001100984
network_acronym_str SBDPC-1
network_name_str Acta Cirúrgica Brasileira (Online)
repository_id_str
spelling Aldefluor protocol to sort keratinocytes stem cells from skinKeratinocytesFlow CytometryAdult Stem Cells.Abstract Purpose: To investigate the use Aldefluor® and N, N - Dimethylaminobenzaldehyde (DEAB) to design a protocol to sort keratinocyte stem cells from cultured keratinocytes from burned patients. Methods: Activated Aldefluor® aliquots were prepared and maintained at temperature between 2 to 8°C, or stored at -20°C. Next, the cells were collected following the standard protocol of sample preparation. Results: Best results were obtained with Aldefluor® 1.5µl and DEAB 15 µl for 1 x 106 cells, incubated at 37°C for 15 minutes. Flow cytometer range for keratinocyte stem cells separation was evaluated. There were 14.8% of stem cells separated in one sample of keratinocyte culture used to pattern the protocol. After being defined the ideal concentration, the same test pattern was performed in other keratinocyte samples. We observed a final mean of 10.8%. Conclusion: Aldefluor® has been shown as a favorable marking of epidermal keratinocyte stem cells for subsequent separation on a flow cytometer, with detection of 10.8% of epidermal keratinocyte stem cells, in this protocol.Sociedade Brasileira para o Desenvolvimento da Pesquisa em Cirurgia2017-11-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0102-86502017001100984Acta Cirúrgica Brasileira v.32 n.11 2017reponame:Acta Cirúrgica Brasileira (Online)instname:Sociedade Brasileira para o Desenvolvimento da Pesquisa em Cirurgia (SBDPC)instacron:SBDPC10.1590/s0102-865020170110000010info:eu-repo/semantics/openAccessNoronha,Samuel Marcos RibeiroGragnani,AlfredoPereira,Thiago Antônio CaladoCorrea,Silvana Aparecida AlvesBonucci,JessicaFerreira,Lydia Masakoeng2017-12-19T00:00:00Zoai:scielo:S0102-86502017001100984Revistahttps://www.bvs-vet.org.br/vetindex/periodicos/acta-cirurgica-brasileira/https://old.scielo.br/oai/scielo-oai.php||sgolden@terra.com.br0102-86501678-2674opendoar:2017-12-19T00:00Acta Cirúrgica Brasileira (Online) - Sociedade Brasileira para o Desenvolvimento da Pesquisa em Cirurgia (SBDPC)false
dc.title.none.fl_str_mv Aldefluor protocol to sort keratinocytes stem cells from skin
title Aldefluor protocol to sort keratinocytes stem cells from skin
spellingShingle Aldefluor protocol to sort keratinocytes stem cells from skin
Noronha,Samuel Marcos Ribeiro
Keratinocytes
Flow Cytometry
Adult Stem Cells.
title_short Aldefluor protocol to sort keratinocytes stem cells from skin
title_full Aldefluor protocol to sort keratinocytes stem cells from skin
title_fullStr Aldefluor protocol to sort keratinocytes stem cells from skin
title_full_unstemmed Aldefluor protocol to sort keratinocytes stem cells from skin
title_sort Aldefluor protocol to sort keratinocytes stem cells from skin
author Noronha,Samuel Marcos Ribeiro
author_facet Noronha,Samuel Marcos Ribeiro
Gragnani,Alfredo
Pereira,Thiago Antônio Calado
Correa,Silvana Aparecida Alves
Bonucci,Jessica
Ferreira,Lydia Masako
author_role author
author2 Gragnani,Alfredo
Pereira,Thiago Antônio Calado
Correa,Silvana Aparecida Alves
Bonucci,Jessica
Ferreira,Lydia Masako
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Noronha,Samuel Marcos Ribeiro
Gragnani,Alfredo
Pereira,Thiago Antônio Calado
Correa,Silvana Aparecida Alves
Bonucci,Jessica
Ferreira,Lydia Masako
dc.subject.por.fl_str_mv Keratinocytes
Flow Cytometry
Adult Stem Cells.
topic Keratinocytes
Flow Cytometry
Adult Stem Cells.
description Abstract Purpose: To investigate the use Aldefluor® and N, N - Dimethylaminobenzaldehyde (DEAB) to design a protocol to sort keratinocyte stem cells from cultured keratinocytes from burned patients. Methods: Activated Aldefluor® aliquots were prepared and maintained at temperature between 2 to 8°C, or stored at -20°C. Next, the cells were collected following the standard protocol of sample preparation. Results: Best results were obtained with Aldefluor® 1.5µl and DEAB 15 µl for 1 x 106 cells, incubated at 37°C for 15 minutes. Flow cytometer range for keratinocyte stem cells separation was evaluated. There were 14.8% of stem cells separated in one sample of keratinocyte culture used to pattern the protocol. After being defined the ideal concentration, the same test pattern was performed in other keratinocyte samples. We observed a final mean of 10.8%. Conclusion: Aldefluor® has been shown as a favorable marking of epidermal keratinocyte stem cells for subsequent separation on a flow cytometer, with detection of 10.8% of epidermal keratinocyte stem cells, in this protocol.
publishDate 2017
dc.date.none.fl_str_mv 2017-11-01
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=S0102-86502017001100984
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0102-86502017001100984
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/s0102-865020170110000010
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 para o Desenvolvimento da Pesquisa em Cirurgia
publisher.none.fl_str_mv Sociedade Brasileira para o Desenvolvimento da Pesquisa em Cirurgia
dc.source.none.fl_str_mv Acta Cirúrgica Brasileira v.32 n.11 2017
reponame:Acta Cirúrgica Brasileira (Online)
instname:Sociedade Brasileira para o Desenvolvimento da Pesquisa em Cirurgia (SBDPC)
instacron:SBDPC
instname_str Sociedade Brasileira para o Desenvolvimento da Pesquisa em Cirurgia (SBDPC)
instacron_str SBDPC
institution SBDPC
reponame_str Acta Cirúrgica Brasileira (Online)
collection Acta Cirúrgica Brasileira (Online)
repository.name.fl_str_mv Acta Cirúrgica Brasileira (Online) - Sociedade Brasileira para o Desenvolvimento da Pesquisa em Cirurgia (SBDPC)
repository.mail.fl_str_mv ||sgolden@terra.com.br
_version_ 1752126444544196608