Single-cell qPCR facilitates the optimization of hematopoietic differentiation in hPSCs/OP9 coculture system
Braz. j. med. biol. res
; 51(5): e7183, 2018. graf
Article
em En
| LILACS
| ID: biblio-889088
Biblioteca responsável:
BR1.1
ABSTRACT
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.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
LILACS
Assunto principal:
Células-Tronco Hematopoéticas
/
Diferenciação Celular
/
Técnicas de Cocultura
/
Células-Tronco Pluripotentes
Limite:
Humans
Idioma:
En
Revista:
Braz. j. med. biol. res
Assunto da revista:
BIOLOGIA
/
MEDICINA
Ano de publicação:
2018
Tipo de documento:
Article
/
Project document
País de afiliação:
China
País de publicação:
Brasil