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Efficient gene transfer of a simian immuno‐deficiency viral vector into cardiomyocytes derived from primate embryonic stem cells
Author(s) -
Nagata Mihoko,
Takahashi Masafumi,
Muramatsu Shinichi,
Ueda Yasuji,
Hanazono Yutaka,
Takeuchi Koichi,
Okada Koji,
Suzuki Yutaka,
Kondo Yasushi,
Suemori Masafumi,
Ikeda Uichi,
Nakano Imaharu,
Kobayashi Eiji,
Hasegawa Mamoru,
Ozawa Keiya,
Nakatsuji Norio,
Shimada Kazuyuki
Publication year - 2003
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.431
Subject(s) - embryonic stem cell , biology , viral vector , microbiology and biotechnology , genetic enhancement , transduction (biophysics) , green fluorescent protein , primate , transfection , stem cell , cell type , immunostaining , gene delivery , cell culture , cell , gene , immunology , genetics , neuroscience , immunohistochemistry , biochemistry , recombinant dna
Background Embryonic stem (ES) cells continually proliferate and can generate large numbers of differentiated cells. Genetic manipulation of transplantable cells derived from primate ES cells offers considerable potential for development research and regenerative cell therapy. However, protocols for efficient gene transfer into primate ES‐cell‐derived cells have not yet been established. Methods Spontaneously contracting areas were derived from cynomolgus monkey ES cells. Features of cardiomyocytes in the area were analyzed according to gene expression (RT‐PCR), morphology (immunostaining and electron microscopy), and function (intracellular calcium transience). Beating cells were transduced using a simian immunodeficiency virus (SIV) vector expressing enhanced green fluorescence protein (EGFP), then transplanted into ischemic rat myocardium. Results Beating cells derived from monkey ES cells displayed gene expression, ultrastructural and functional properties of early‐stage cardiomyocytes. Highly efficient (97% cardiac phenotype) and stable transduction of these ES‐cell‐derived cardiomyocytes was achieved using SIV vector without altering contractile function. In addition, transduced cardiomyocytes survived in the myocardium of a rat myocardial infarction model. Conclusions A lentiviral vector system based on SIV represents a useful vehicle for genetic modification of cardiomyocytes derived from primate ES cells, and can extend the application of primate ES cells to gene therapy. Copyright © 2003 John Wiley & Sons, Ltd.

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