Single-cell qPCR facilitates the optimization of hematopoietic differentiation in hPSCs/OP9 coculture system

Detalhes bibliográficos
Autor(a) principal: Chen,Haide
Data de Publicação: 2018
Outros Autores: Jiang,Mengmeng, Xiao,Lei, Huang,He
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Journal of Medical and Biological Research
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2018000500608
Resumo: Human pluripotent stem cells (hPSCs)/OP9 coculture system is a widely used hematopoietic differentiation approach. The limited understanding of this process leads to its low efficiency. Thus, we used single-cell qPCR to reveal the gene expression profiles of individual CD34+ cells from different stages of differentiation. According to the dynamic gene expression of hematopoietic transcription factors, we overexpressed specific hematopoietic transcription factors (Gata2, Lmo2, Etv2, ERG, and SCL) at an early stage of hematopoietic differentiation. After overexpression, we generated more CD34+ cells with normal expression level of CD43 and CD31, which are used to define various hematopoietic progenitors. Furthermore, these CD34+ cells possessed normal differentiation potency in colony-forming unit assays and normal gene expression profiles. In this study, we demonstrated that single-cell qPCR can provide guidance for optimization of hematopoietic differentiation and transient overexpression of selected hematopoietic transcription factors can enhance hematopoietic differentiation.
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spelling Single-cell qPCR facilitates the optimization of hematopoietic differentiation in hPSCs/OP9 coculture systemSingle-cell qPCRHuman pluripotent stem cellsHematopoietic differentiationOP9-GFPTranscription factorsHuman pluripotent stem cells (hPSCs)/OP9 coculture system is a widely used hematopoietic differentiation approach. The limited understanding of this process leads to its low efficiency. Thus, we used single-cell qPCR to reveal the gene expression profiles of individual CD34+ cells from different stages of differentiation. According to the dynamic gene expression of hematopoietic transcription factors, we overexpressed specific hematopoietic transcription factors (Gata2, Lmo2, Etv2, ERG, and SCL) at an early stage of hematopoietic differentiation. After overexpression, we generated more CD34+ cells with normal expression level of CD43 and CD31, which are used to define various hematopoietic progenitors. Furthermore, these CD34+ cells possessed normal differentiation potency in colony-forming unit assays and normal gene expression profiles. In this study, we demonstrated that single-cell qPCR can provide guidance for optimization of hematopoietic differentiation and transient overexpression of selected hematopoietic transcription factors can enhance hematopoietic differentiation.Associação Brasileira de Divulgação Científica2018-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2018000500608Brazilian Journal of Medical and Biological Research v.51 n.5 2018reponame:Brazilian Journal of Medical and Biological Researchinstname:Associação Brasileira de Divulgação Científica (ABDC)instacron:ABDC10.1590/1414-431x20187183info:eu-repo/semantics/openAccessChen,HaideJiang,MengmengXiao,LeiHuang,Heeng2019-03-19T00:00:00Zoai:scielo:S0100-879X2018000500608Revistahttps://www.bjournal.org/https://old.scielo.br/oai/scielo-oai.phpbjournal@terra.com.br||bjournal@terra.com.br1414-431X0100-879Xopendoar:2019-03-19T00:00Brazilian Journal of Medical and Biological Research - Associação Brasileira de Divulgação Científica (ABDC)false
dc.title.none.fl_str_mv Single-cell qPCR facilitates the optimization of hematopoietic differentiation in hPSCs/OP9 coculture system
title Single-cell qPCR facilitates the optimization of hematopoietic differentiation in hPSCs/OP9 coculture system
spellingShingle Single-cell qPCR facilitates the optimization of hematopoietic differentiation in hPSCs/OP9 coculture system
Chen,Haide
Single-cell qPCR
Human pluripotent stem cells
Hematopoietic differentiation
OP9-GFP
Transcription factors
title_short Single-cell qPCR facilitates the optimization of hematopoietic differentiation in hPSCs/OP9 coculture system
title_full Single-cell qPCR facilitates the optimization of hematopoietic differentiation in hPSCs/OP9 coculture system
title_fullStr Single-cell qPCR facilitates the optimization of hematopoietic differentiation in hPSCs/OP9 coculture system
title_full_unstemmed Single-cell qPCR facilitates the optimization of hematopoietic differentiation in hPSCs/OP9 coculture system
title_sort Single-cell qPCR facilitates the optimization of hematopoietic differentiation in hPSCs/OP9 coculture system
author Chen,Haide
author_facet Chen,Haide
Jiang,Mengmeng
Xiao,Lei
Huang,He
author_role author
author2 Jiang,Mengmeng
Xiao,Lei
Huang,He
author2_role author
author
author
dc.contributor.author.fl_str_mv Chen,Haide
Jiang,Mengmeng
Xiao,Lei
Huang,He
dc.subject.por.fl_str_mv Single-cell qPCR
Human pluripotent stem cells
Hematopoietic differentiation
OP9-GFP
Transcription factors
topic Single-cell qPCR
Human pluripotent stem cells
Hematopoietic differentiation
OP9-GFP
Transcription factors
description Human pluripotent stem cells (hPSCs)/OP9 coculture system is a widely used hematopoietic differentiation approach. The limited understanding of this process leads to its low efficiency. Thus, we used single-cell qPCR to reveal the gene expression profiles of individual CD34+ cells from different stages of differentiation. According to the dynamic gene expression of hematopoietic transcription factors, we overexpressed specific hematopoietic transcription factors (Gata2, Lmo2, Etv2, ERG, and SCL) at an early stage of hematopoietic differentiation. After overexpression, we generated more CD34+ cells with normal expression level of CD43 and CD31, which are used to define various hematopoietic progenitors. Furthermore, these CD34+ cells possessed normal differentiation potency in colony-forming unit assays and normal gene expression profiles. In this study, we demonstrated that single-cell qPCR can provide guidance for optimization of hematopoietic differentiation and transient overexpression of selected hematopoietic transcription factors can enhance hematopoietic differentiation.
publishDate 2018
dc.date.none.fl_str_mv 2018-01-01
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=S0100-879X2018000500608
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2018000500608
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/1414-431x20187183
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 Associação Brasileira de Divulgação Científica
publisher.none.fl_str_mv Associação Brasileira de Divulgação Científica
dc.source.none.fl_str_mv Brazilian Journal of Medical and Biological Research v.51 n.5 2018
reponame:Brazilian Journal of Medical and Biological Research
instname:Associação Brasileira de Divulgação Científica (ABDC)
instacron:ABDC
instname_str Associação Brasileira de Divulgação Científica (ABDC)
instacron_str ABDC
institution ABDC
reponame_str Brazilian Journal of Medical and Biological Research
collection Brazilian Journal of Medical and Biological Research
repository.name.fl_str_mv Brazilian Journal of Medical and Biological Research - Associação Brasileira de Divulgação Científica (ABDC)
repository.mail.fl_str_mv bjournal@terra.com.br||bjournal@terra.com.br
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