Aldefluor protocol to sort keratinocytes stem cells from skin
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Outros Autores: | , , , , |
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. |
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Acta Cirúrgica Brasileira (Online) |
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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 |