1996 Fiscal Year Final Research Report Summary
Establishment of cardiac myocyte cell line
| Project/Area Number |
06557042
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | UNIVERSITY OF TOKYO |
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Principal Investigator |
KOMURO Issei University of Tokyo, Third Depertment of Internal Medicine. Instructor, 医学部・附属病院, 助手 (30260483)
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| Co-Investigator(Kenkyū-buntansha) |
KATOH Hirohisa Yamasa Shouyu Inc. Department of Developmental Rescarch. rescatcher, 研究開発部, 研究員
KUROO Makoto National Institute of Neuroscience Division of Molecular Genetics. researcher, 研究員
YAMAZAKI Tsutomu University of Tokyo, Health Service Center. Assistant Professor, 保健センター, 講師 (60251245)
YAZAKI Yoshio University of Tokyo, Third Depertment of Internal Medicine. Professor, 医学部・附属病院, 教授 (20101090)
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| Project Period (FY) |
1994 – 1996
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| Keywords | embrvonic cell / transcription factor / cardiac development / MAP kinase / homeobox / zine finger / tumer suppresser gene |
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| Research Abstract |
We have established cardiac differentiation system using embrryonic stem (ES) cells. When ES cells were cultured in suspension, they developed to embryoid bodies (EB) and started to beat spontaneously from 8 days. Northern blot analysis revealed that cardiac specific homeoprotein CSX and zinc finger protein GATA4 were expressed in EB from 5 days and contractile protein genes such as alpha myosin heavy chain and myosin light chain genes were expressed from 8 days after initiation of suspension culture. We examined the role of mitogen-activated protein kinase (MAPK) in cardiac differentiation using this system. Dominant negative MAPK strongly suppressed transcription of CSX and constitutively activated MAPK conversely activated transcription of CSX.We next isolated ES cell lines (designated as ES-MKP) which were permanently transfected by MAPK phosphatase (MKP), which is a negative regulator of MAPK.When ES-MKP was cultured in suspension, a few EB showed spontaneous contraction and expressed a little myosin heavy chain protein. These results suggest that MAPK is necessary for the development of cardiac myocytes.
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