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
Dans Anglais
| LILACS
| ID: biblio-889088
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.
Texte intégral:
Disponible
Indice:
LILAS (Amériques)
Sujet Principal:
Cellules souches hématopoïétiques
/
Différenciation cellulaire
/
Techniques de coculture
/
Cellules souches pluripotentes
Limites du sujet:
Humains
langue:
Anglais
Texte intégral:
Braz. j. med. biol. res
Thème du journal:
Biologie
/
Médicament
Année:
2018
Type:
Article
/
descriptif de projet
Pays d'affiliation:
Chine
Institution/Pays d'affiliation:
Zhejiang University/CN
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