Cardiomyocytes can be generated from marrow stromal cells in vitro

Shinji Makino, Keiichi Fukuda, Shunichirou Miyoshi, Fusako Konishi, Hiroaki Kodama, Jing Pan, Motoaki Sano, Toshiyuki Takahashi, Shingo Hori, Hitoshi Abe, Jun Ichi Hata, Akihiro Umezawa, Satoshi Ogawa

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1644 Citations (Scopus)

Abstract

We have isolated a cardiomyogenic cell line (CMG) from murine bone marrow stromal cells. Stromal cells were immortalized, treated with 5- azacytidine, and spontaneously beating cells were repeatedly screened. The cells showed a fibroblast-like morphology, but the morphology changed after 5-azacytidine treatment in -30% of the cells; they connected with adjoining cells after one week, formed myotube-like structures, began spontaneously beating after two weeks, and beat synchronously after three weeks. They expressed atrial natriuretic peptide and brain natriuretic peptide and were stained with anti-myosin, anti-desmin, and anti-actinin antibodies. Electron microscopy revealed a cardiomyocyte-like ultrastructure, including typical sarcomeres, a centrally positioned nucleus, and atrial granules. These cells had several types of action potentials, such as sinus node-like and ventricular cell-like action potentials. All cells had a long action potential duration or plateau, a relatively shallow resting membrane potential, and a pacemaker-like late diastolic slow depolarization. Analysis of the isoform of contractile protein genes, such as myosin heavy chain, myosin light chain, and α-actin, indicated that their muscle phenotype was similar to that of fetal ventricular cardiomyocytes. These cells expressed Nkx2.5/Csx, GATA4, TEF-1, and MEF-2C mRNA before 5-azacytidine treatment and expressed MEF-2A and MEF-2D after treatment. This new cell line provides a powerful model for the study of cardiomyocyte differentiation.

Original languageEnglish
Pages (from-to)697-705
Number of pages9
JournalJournal of Clinical Investigation
Volume103
Issue number5
DOIs
Publication statusPublished - 1999 Mar

ASJC Scopus subject areas

  • Medicine(all)

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    Makino, S., Fukuda, K., Miyoshi, S., Konishi, F., Kodama, H., Pan, J., Sano, M., Takahashi, T., Hori, S., Abe, H., Hata, J. I., Umezawa, A., & Ogawa, S. (1999). Cardiomyocytes can be generated from marrow stromal cells in vitro. Journal of Clinical Investigation, 103(5), 697-705. https://doi.org/10.1172/JCI5298