Cargando…

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

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 s...

Descripción completa

Detalles Bibliográficos
Autores principales: Chen, Haide, Jiang, Mengmeng, Xiao, Lei, Huang, He
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Associação Brasileira de Divulgação Científica 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5875903/
https://www.ncbi.nlm.nih.gov/pubmed/29561959
http://dx.doi.org/10.1590/1414-431X20187183
_version_ 1783310438166953984
author Chen, Haide
Jiang, Mengmeng
Xiao, Lei
Huang, He
author_facet Chen, Haide
Jiang, Mengmeng
Xiao, Lei
Huang, He
author_sort Chen, Haide
collection PubMed
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.
format Online
Article
Text
id pubmed-5875903
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher Associação Brasileira de Divulgação Científica
record_format MEDLINE/PubMed
spelling pubmed-58759032018-04-06 Single-cell qPCR facilitates the optimization of hematopoietic differentiation in hPSCs/OP9 coculture system Chen, Haide Jiang, Mengmeng Xiao, Lei Huang, He Braz J Med Biol Res Research Articles 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. Associação Brasileira de Divulgação Científica 2018-03-15 /pmc/articles/PMC5875903/ /pubmed/29561959 http://dx.doi.org/10.1590/1414-431X20187183 Text en https://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Chen, Haide
Jiang, Mengmeng
Xiao, Lei
Huang, He
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
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_short 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
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5875903/
https://www.ncbi.nlm.nih.gov/pubmed/29561959
http://dx.doi.org/10.1590/1414-431X20187183
work_keys_str_mv AT chenhaide singlecellqpcrfacilitatestheoptimizationofhematopoieticdifferentiationinhpscsop9coculturesystem
AT jiangmengmeng singlecellqpcrfacilitatestheoptimizationofhematopoieticdifferentiationinhpscsop9coculturesystem
AT xiaolei singlecellqpcrfacilitatestheoptimizationofhematopoieticdifferentiationinhpscsop9coculturesystem
AT huanghe singlecellqpcrfacilitatestheoptimizationofhematopoieticdifferentiationinhpscsop9coculturesystem