Development of regenerative cardiomyocytes from mesenchymal stem cells for cardiovascular tissue engineering.
We have isolated a cardiomyogenic (CMG) cell line from murine bone marrow stroma. Stromal cells were immortalized, treated with 5-azacytidine, and spontaneous beating cells were repeatedly screened for. The cells showed a fibroblast-like morphology. However, this morphology changed after 5-azacytidine treatment in about 30% of the cells, which connected with adjoining cells after 1 week, formed myotube-like structures and began spontaneous beating after 2 weeks, and beat synchronously after 3 weeks. These cells expressed atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP). Electron microscopy revealed a cardiomyocyte-like ultrastructure including typical sarcomeres and atrial granules. They had sinus node-like or ventricular cell-like action potentials. Analysis of the isoform of contractile protein genes, such as myosin and alpha-actin, indicated that their phenotype was similar to fetal ventricular cardiomyocytes. These cells expressed Nkx2.5, GATA4, TEF-1, and MEF2-C mRNA before 5-azacytidine treatment, and expressed MEF2-A and MEF2-D after treatment. This new cell line provides a powerful model for the study of cardiomyocyte transplantation.
Fukuda
K
K
Institute for Advanced Cardiac Therapeutics, Keio University School of Medicine, Tokyo, Japan. kfukuda@mc.med.keio.ac.jp
D000199:Actins / Q000032:analysis; D000200:Action Potentials; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000032:analysis; D001374:Azacitidine / Q000494:pharmacology; D001698:Biomedical Engineering; D001854:Bone Marrow Cells; D002454:Cell Differentiation; D002460:Cell Line; D017690:Cell Transplantation; D005260:Female; D006321:Heart / Q000502:physiology; D007150:Immunohistochemistry; D008648:Mesoderm / Q000166:cytology; D051379:Mice; D008809:Mice, Inbred C3H; D008854:Microscopy, Electron; D009200:Myocardial Contraction; D009206:Myocardium / Q000737:chemistry / Q000166:cytology* / Q000648:ultrastructure; D009218:Myosins / Q000032:analysis; D020097:Natriuretic Peptide, Brain / Q000032:analysis; D012038:Regeneration; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology*; D017154:Stromal Cells / Q000166:cytology
D000199:Actins; D020097:Natriuretic Peptide, Brain; D009320:Atrial Natriuretic Factor; D009218:Myosins; D001374:Azacitidine