Efficient generation of human embryonic stem cell-derived cardiac progenitors based on tissue-specific enhanced green fluorescence protein expression

Kornélia Szebényi, Adrienn Péntek, Zsuzsa Erdei, György Várady, Tamás I. Orbán, B. Sarkadi, A. Apáti

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Cardiac progenitor cells (CPCs) are committed to the cardiac lineage but retain their proliferative capacity before becoming quiescent mature cardiomyocytes (CMs). In medical therapy and research, the use of human pluripotent stem cell-derived CPCs would have several advantages compared with mature CMs, as the progenitors show better engraftment into existing heart tissues, and provide unique potential for cardiovascular developmental as well as for pharmacological studies. Here, we demonstrate that the CAG promoter-driven enhanced green fluorescence protein (EGFP) reporter system enables the identification and isolation of embryonic stem cell-derived CPCs. Tracing of CPCs during differentiation confirmed up-regulation of surface markers, previously described to identify cardiac precursors and early CMs. Isolated CPCs express cardiac lineage-specific transcripts, still have proliferating capacity, and can be re-aggregated into embryoid body-like structures (CAG-EGFPhigh rEBs). Expression of troponin T and NKX2.5 mRNA is up-regulated in long-term cultured CAG-EGFPhigh rEBs, in which more than 90% of the cells become Troponin I positive mature CMs. Moreover, about one third of the CAG-EGFPhigh rEBs show spontaneous contractions. The method described here provides a powerful tool to generate expandable cultures of pure human CPCs that can be used for exploring early markers of the cardiac lineage, as well as for drug screening or tissue engineering applications.

Original languageEnglish
Pages (from-to)35-45
Number of pages11
JournalTissue Engineering - Part C: Methods
Volume21
Issue number1
DOIs
Publication statusPublished - Jan 1 2015

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Stem cells
Stem Cells
Fluorescence
Tissue
Proteins
Cardiac Myocytes
Troponin T
Troponin I
Tissue engineering
Cell culture
Screening
Messenger RNA
Pharmaceutical Preparations
Embryoid Bodies
Pluripotent Stem Cells
Preclinical Drug Evaluations
Tissue Engineering
Embryonic Stem Cells
Human Embryonic Stem Cells
Biomedical Research

ASJC Scopus subject areas

  • Biomedical Engineering
  • Bioengineering
  • Medicine (miscellaneous)

Cite this

Efficient generation of human embryonic stem cell-derived cardiac progenitors based on tissue-specific enhanced green fluorescence protein expression. / Szebényi, Kornélia; Péntek, Adrienn; Erdei, Zsuzsa; Várady, György; Orbán, Tamás I.; Sarkadi, B.; Apáti, A.

In: Tissue Engineering - Part C: Methods, Vol. 21, No. 1, 01.01.2015, p. 35-45.

Research output: Contribution to journalArticle

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