Enhanced Differentiation of Human Pluripotent Stem Cells into Cardiomyocytes by Bacteria-mediated Transcription Factors Delivery

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2018 Mar 27

PMID 29579079

Citations 6

Authors

Yongxin Jin

Ying Liu

Zhenpeng Li

Katherine Santostefano

Jing Shi

Xinwen Zhang

Donghai Wu

Zhihui Cheng

Weihui Wu

Naohiro Terada

Shouguang Jin

Fang Bai

Affiliations

Soon will be listed here.

Abstract

Virus-mediated expression of defined transcription factor (TF) genes can effectively induce cellular reprogramming. However, sustained expression of the TFs often hinders pluripotent stem cell (PSC) differentiation into specific cell types, as each TF exerts its effect on PSCs for a defined period of time during differentiation. Here, we applied a bacterial type III secretion system (T3SS)-based protein delivery tool to directly translocate TFs in the form of protein into human PSCs. This transient protein delivery technique showed high delivery efficiency for hPSCs, and it avoids potential genetic alterations caused by the introduction of transgenes. In an established cardiomyocyte de novo differentiation procedure, five transcriptional factors, namely GATA4, MEF2C, TBX5, ESRRG and MESP1 (abbreviated as GMTEM), were translocated at various time points. By detecting the expression of cardiac marker genes (Nkx2.5 and cTnT), we found that GMTEM proteins delivered on mesoderm stage of the cardiomyocytes lineage differentiation significantly enhanced both the human ESC and iPSC differentiation into cardiomyocytes, while earlier or later delivery diminished the enhancing effect. Furthermore, all of the five factors were required to enhance the cardiac differentiation. This work provides a virus-free strategy of transient transcription factors delivery for directing human stem cell fate without jeopardizing genome integrity, thus safe for biomedical applications.

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