| Project/Area Number |
13671401
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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 |
Thoracic surgery
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| Research Institution | NARA MEDICAL UNIVERSITY |
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Principal Investigator |
TANIGUCHI Shigeki Nara Medical University, Surgery III, Professor, 第三外科, 教授 (90183467)
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| Co-Investigator(Kenkyū-buntansha) |
NAITO Hiroshi Nara Medical University, Surgery III, Research Associate, 第三外科, 助手
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2002: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | embryonic stem cell / cardiomyocyte / cell transplantation / myocardial infarction / 移植 |
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| Research Abstract |
The purpose of this study was to investigate the survival of embryonic stem cell (ES cell)-derived cardiomyocytes transplanted into the infarcted myocardium.
In in vitro study, undifferentiated mouse ES cells carrying the enhanced green fluorescent protein (EGFP) were cultured in hanging drops for 7 days and plated onto dishes, and then beating clusters were dissected and used for the following analyses and transplantation. These cells were identified as cardiomyocytes by reverse transcription-polymerase chain reaction of cardiac-specific genes, recording of action potential, and immunostaining of sarcomeric myosin staining. As a result of in vitro study, ES cell-derived cardiomyocytes were shown to have myosin light chain-2v and alpha-myosin heavy chain genes expression, action potential like one from a sinus nodal cell and a ventricular myocyte, and positive for sarcomeric myosin staining.
In in vivo study, a myocardial infarction rat model was made by the ligation of the left coronary artery. Beating clusters were injected into the border zone between the infarcted and normal myocardium. Ten and 30 days after transplantation EGFP expression, hematoxylin-eosin staining and immunostaining of sarcomeric myosin were investigated. As a result of in vivo study, EGFP-expressed cells were detected after transplantation, and a few of them were positive for sarcomeric myosin. No lymphocytic infiltration was observed.
These results suggested that ES cells differentiated into cardiomyocytes in vitro and ES cell-derived cardiomyocytes could be transplanted into the infarcted myocardium and survive in vivo.
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