Phenotype-specific cells with proliferative potential are produced by polyethylene glycol-induced fusion of mouse embryonic stem cells with fetal cardiomyocytes.
Because cardiomyocytes lose the ability to divide upon differentiation, myocardial failure is assumed to be generally irreversible. For terminal cardiac insufficiency, the potential for regenerative treatment by stem cells, especially embryonic stem (ES) cells, offers hope for the future. Recent studies showed that stem cells fuse spontaneously with cells remaining in damaged tissues, and restore tissue function. To imitate spontaneous fusion in vivo, we used polyethylene glycol (PEG) in vitro to fuse mouse ES cells and fetal cardiomyocytes and analyzed the cytochemical properties of the fused cells. Confocal laser scanning microscopy coupled with lipophilic dye labeling of the living cell membranes showed that there were fused cells of ES cells and cardiomyocytes after PEG treatment. By flow cytometry, the fusion efficiency between ES cells and cardiomyocytes was estimated to be about 45% of the total resulting cells. When green fluorescent protein (GFP)-expressing ES cells were fused with cardiomyocytes, the fused cells had immunoreactivity for GFP in their cytoplasm and cardiac troponin I in their myofibrils. Some of these cells also expressed proliferating cell nuclear antigen up to 11 days after fusion, the last time point examined. This study shows that PEG-induced fusions of mouse ES cells and cardiomyocytes have the cardiomyocyte phenotype and proliferation potential.
Takei
Shunsuke
S
Department of Anatomy and Organ Technology, Shinshu University School of Medicine, Matsumoto, Japan. takei@sch.md.shinshu-u.ac.jp
Yamamoto
Makoto
M
Cui
Li
L
Yue
Fengming
F
Johkura
Kohei
K
Ogiwara
Naoko
N
Iinuma
Hisae
H
Okinaga
Kota
K
Sasaki
Katsunori
K
D000818:Animals; D002459:Cell Fusion / Q000379:methods*; D048708:Cell Growth Processes / Q000502:physiology; D005260:Female; D005318:Fetal Heart / Q000166:cytology*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011247:Pregnancy; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
D049452:Green Fluorescent Proteins; D011092:Polyethylene Glycols