| Project/Area Number |
15609007
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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 |
幹細胞生物学
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| Research Institution | National Cardiovascular Center Research Institute |
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Principal Investigator |
HIDAKA Kyoko National Cardiovascular Center Research Institute, Department of Bioscience, Laboratory Chief, バイオサイエンス部, 室長 (00216681)
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| Co-Investigator(Kenkyū-buntansha) |
MORISAKI Takayuki National Cardiovascular Center Research Institute, Department of Bioscience, Director, バイオサイエンス部, 部長 (30174410)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2003: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | ES Cells / Cardiomyocytes / Nkx2-5 / ErbB4 / Wnt11 / 心筋分化誘導 / ErbB / Nkx2.5 |
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| Research Abstract |
Although ES cells spontaneously differentiate into various cell types, only a part of cells differentiate into cardiomyocytes. We have established the cell line, in which the EGFP gene was knocked into the locus of the cardiac homeobox transcription factor gene, Nkx2-5. Using the cell line, we characterized the ES cell-derived cardiomyocytes and searched for methods to induce and isolate cardiomyocytes derived from ES cells.
1.We have characterized Nkx2-5/EGFP-positive cells derived from ES cells, and found that they represented cardiac progenitor cells able to differentiate ventricular, atrial or conduction system cells. We also analyzed Ca2+ channels expressed in Nkx2-5/EGFP-positive cells.
2.The ErbB4 gene was found to be preferentially expressed in the Nkx2-5/EGFP cells. Antagonists for ErbB signaling inhibited cardiogenesis of ES cells. Administration of neuregulin, a ligand of ErbB, increased (1.5 fold) the population of cardiomyocytes derived from ES cells.
3.The Wnt11 gene was expressed in the spatio-temporally similar pattern with Nkx2-5. Administration of Wnt11-conditioned medium increased (1.5 fold) the population of cardiomyocytes derived from ES cells.
4.Since the promoter of Nkx2-5 was not so strong, we searched other cardiac-specific promoters to drive the reporter genes. The myosin heavy chain alpha gene promoter was found to be suitable to isolate cardiomyotyes derived from ES cells.
5.The Luc71 gene, preferentially expressed in undifferentiated stem cells, was found to be important for the differentiation of myocytes.
Thus, we found that several signaling pathways including ErbB4 and Wnt11 were important to induce cardiomyocytes from ES cells. We also improved methods to isolate cardiomyocytes by choosing strong promoters. These findings will be valuable to develop efficient methods to isolate cardiomyocytes from ES cells for regenerative medicine.
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