Isolation and characterization of skeletal muscle-derived cardiac progenitor cells for cardiac regeneration therapy

2004 Fiscal Year Final Research Report Summary

• Project/Area Number: 15590720
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Circulatory organs internal medicine
• Research Institution: Chiba University
• Principal Investigator: NAGAI Toshio Chiba University, Univ.Hospital, Assistant Professor, 医学部附属病院, 講師 (00334194)
• Co-Investigator(Kenkyū-buntansha): KOMURO Issei Chiba University, Graduate School of Medicine, Professor, 大学院・医学研究院, 教授 (30260483)
• Project Period (FY): 2003 – 2004
• Keywords: skeletal muscle / stem cells / transdifferentiation / cell fusion / cardiomyocytes

Research Abstract

When green fluorescent proteins(GFP) positive skeletal muscle cells were co-cultured with cardiomyocytes some of GFP positive cells expressed cardiac specific troponinT (cTnT), suggesting that skeletal muscle derived cells transdifferentiate into cardiomyocytes. In the first year we examined whether mechanical stress, Ca2+, growth factors, calmodulin and calcineurin effect on the process of transdifferentiation. When the co-cultured cells were treated with niffedipine, calmodulin inhibitors or calcineurin inhibitors the number of cTnT positive skeletal muscle cells decreased. Mechanical stress attenuated the inhibitory effect of niffedipine. In the second year we examined the mechanisms through which skeletal muscle cells acquire the phenotypes of cardiomyocytes by using the co-culture method. When skeletal muscle cells derived from GFP transgenic mouse were co-cultured with cardiomyocytes expressing LacZ gene 70-80% of GFP and cTnT double positive cells expressed b-galacfocidase, sugg esting that cell fusion took place. We next examined whether other somatic cells fuse with cardiomyocytes and maintain cardiac phenotypes. When cardiomyocytes were co-cultured with human umbilical vascular endothelial cells(HUVEC) or cardiac fibroblasts some of the cells acquired cardiac phenotypes through cell fusion. During co-culture the ratio of cTnT positive fused cells to the total fused cells increased in time dependent manner. However, the phenotypes of endothelial cells or fibtroblasts in the fused cells were suppressed. When skeletal muscle cells or HUVEC were transplanted into the heart of immunosuppressed rats the fused cells existed in the heart and they expressed cTnT Next we examined whether skeletal muscle-derived stem cells such as side population(SP) cells or Sca-1 positive cells can differentiate into cardiomyocytes. When muscle-derived SP or Sca-1 positive cells were co-cultured with cardiomyocytes the cells acquire the cardiac phenotypes through both fusion and transdifierentiation. However, mono-cultured SP cells or Sca-1 positive cells with various growth factors did not differentiate into cardiomyocytes.

Research Products

• [Journal Article] G-CSF prevents cardiac Remodeling after myocardial infarction by activating Jak/Stat in cardiomyocytes. (2005)
  • Author(s): Harada, M., Qin, Y., Takano, H., Minamino, T., Zou, Y., Toko, H., Ohtsuka, M., Matsuura, K., Sano, M., Nishi, J., Akazawa, H., Kunieda, T., Zhu, W., Hasegawa, H., Kunisada, K., Nagai, T., Nakaya, H., Yamauchi-Takihara, K., Komuro, I.
  • Journal Title: Nat Med Pages: 305-311
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cardiomyocytes fuse with surrounding noncardiomyocytes and reenter the cell cycle (2004)
  • Author(s): Matsuura K
  • Journal Title: J Cell Biol 167 Pages: 351
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Adult cardiac Sca-1 positive cells differentiate into beating cardiomyocytes. (2004)
  • Author(s): Matsuura K
  • Journal Title: J Biol Chem 279 Pages: 11384-11391
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A novel LIM protein Cal promotes cardiac differentiation by association with CSX/NKX2-5. (2004)
  • Author(s): Akazawa H
  • Journal Title: J Cell Biol 164 Pages: 395-405
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] G-CSF prevents cardiac Remodeling after myocardial infarction by activating Jak/Stat in cardiomyocytes. (2004)
  • Author(s): Harada M
  • Journal Title: Nature Medicine 11 Pages: 305-311
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Adult cardiac Sea-1 positive cells differentiate into beating cardiomyocytes. (2004)
  • Author(s): Matsuura, K., Nagai, T., Nishigaki, N., Oyama, T., Nishi, J., Wada, H., Sano, M., Toko, H., Akazawa, H., Sato, T., Nakaya, H, Kasanuki., H., Komuro, I.
  • Journal Title: J Biol Chem 279 Pages: 11384-11391
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Angiotensin II Type la Receptor Is Involved in Cell Infiltration, Cytokine Production, and Neovascularization in Infarcted Myocardium. (2004)
  • Author(s): Toko, H., Zou, Y., Minamino, T., Sakamoto, M., Sano, M., Harada, M., Nagai, T., Sugaya, T., Terasaki, F., Kitaura, Y., Komuro, I.
  • Journal Title: Arterioscler Thromb Vase Biol 24 Pages: 664-670
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A novel LIM protein Cal promotes cardiac differentiation by association with CSX/NKX2-5. (2004)
  • Author(s): Akazawa, H., Kudoh, S., Mochizuki, N., Takekoshi, N., Takano, H., Nagai, T., Komuro, I.
  • Journal Title: J Cell Biol 164 Pages: 395-405
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Diphtheria toxin-induced autophagic cardiomyocyte death plays a pathogenic role in mouse model of heart failure. (2004)
  • Author(s): Akazawa, H., Komazaki, S., Shimomura, H., Terasaki, F., Zou, Y., Takano, H., Nagai, T., Komuro I.
  • Journal Title: J Biol Chem 279 Pages: 41095-41103
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cdc42 plays a critical role in assembly of sarcomere units series of cardiac myocytes. (2003)
  • Author(s): Nagai T
  • Journal Title: Biochem Biophys Res Commun 305 Pages: 806-810
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Beating is necessary for transdifferentiation of skeleteal muscle-derived cells into cardiomyocytes. (2003)
  • Author(s): Iijima Y
  • Journal Title: FASEB J 17 Pages: 1361-1363
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Beating is necessary for transdiffentiation of skeletal muscle-derived cells into cardiomyocytes (2003)
  • Author(s): Iijima, Y., Nagai, T., Mizukami, M., Matsuura, K., Ogura, T., Wada, T., Akazawa, H., Takano, H., Nakaya, H., Komuro, I.
  • Journal Title: FASEB J 17 Pages: 1361-1363
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Stretch-modulation of second messengers : effects on cardiomyocyte ion transport. (2003)
  • Author(s): Kudoh, S., Akazawa, H., Takano, H., Zou, Y., Toko, H., Nagai, T, Komuro, I.
  • Journal Title: Prog Biophys Mol Biol 82 Pages: 57-66
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cdc42 plays a critical role in assembly of sarcomere units series of cardiac myocytes. (2003)
  • Author(s): Nagai, T., Tanaka-Ishikawa, M., Aikawa, R., Ishihara, H., Zhu, W., Yazaki, Y, Nagai, R., Komuro I.
  • Journal Title: Biochem Biophys Res Commun 305 Pages: 806-810
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The role of PPAR γ-dependent pathway in the development of cardiac hypertrophy. (2003)
  • Author(s): Takano, H., Hasegawa, H., Nagai, Tand Komuro, I.
  • Journal Title: Drugs of Today 39 Pages: 347-357
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors prevent the development of cardiac hypertrophy and heart failure in rats. (2003)
  • Author(s): Hasegawa, H., Yamamoto, R., Takano, H., Mizukami, M., Asakawa, M., Nagai, T.Komuro, I.
  • Journal Title: J Mol Cell Cardiol 35 Pages: 923-960
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Heat shock transcription factor 1 protects cardiomyocytes from ischemia/reperfusion injury. (2003)
  • Author(s): Zou, Y., Zhu, W., Sakamoto, M., Qin, Y., Akazawa, H., Toko, H., Mizukami, M., Takeda, N., Minamino, T., Takano, H., Nagai, T., Nakai, A., Komuro, I.
  • Journal Title: Circulation 108 Pages: 3024-3030
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] 冠動脈の臨床(上)-21世紀の診断治療体系-II (2003)
  • Author(s): 永井 敏雄
  • Total Pages: 5
  • Description: 「研究成果報告書概要(和文)」より