| Project/Area Number |
16500270
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neurophysiology and muscle physiology
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| Research Institution | Tokyo Women's Medical University |
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Principal Investigator |
HARAGUCHI Yuji Tokyo Women's Medical University, School of Medicine, Research assistant professor, 医学部, 助手 (80272251)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMIZU Tatsuya Tokyo Women's Medical University, School of Medicine, assistant professor, 医学部, 講師 (40318100)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2005: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | Cell sheet / Temperature-responsive dish / Electrical coupling / Gap junction / Connexin / 心筋細胞シート |
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| Research Abstract |
Previously, we have successfully created pulsatile myocardial tissue grafts using our novel technology, "cell sheet engineering", that layers cell sheets fabricated on temperature-responsive culture dishes to form three-dimensional structures. Electrical coupling is established between layered neonatal rat cardiomyocyte sheets, resulting in the synchronized beating of three-dimensional myocardial tissues. However, the mechanism by which these layered cardiomyocyte sheets communicate electrically is not well understood. In this study, we used a multiple-electrode extracellular recording system and demonstrated that bilayer cardiomyocyte sheets coupled electrically 34 ± 2 minutes (n=24, mean ± SEM) after layering. Immunohistological analysis showed that connexin 43, a gap junction-related protein, existed not only at cell-to-cell interfaces but also on the free cell membrane of the cardiomyocyte sheet. Connexin 43 was detected between bilayer cardiomyocyte sheets at 60 minutes after layering. Dye transfer assay demonstrated that the exchange of small molecules via gap junctions occurred within 30 minutes. These results indicate that direct communication between cardiomyocyte sheets with preserved connexin 43 expression allows for rapid electrical coupling via functional gap junction formation. This is new insight in tissue biology and electrophysiology as well as in myocardial tissue engineering.
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