| Project/Area Number |
06808074
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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 |
Cell biology
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| Research Institution | Tokai University |
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Principal Investigator |
NAKATSUJI Takako Tokai University ・ Institute of Civilization, Associate Professor, 文明研究所(海洋学部), 助教授 (00172346)
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| Co-Investigator(Kenkyū-buntansha) |
NAKATSUJU Norio National Institure of Genetics, Genetic Strains Research Center, Professor., 系統生物研究センター, 教授 (80237312)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1996: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1995: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1994: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | hematopoietic stem cell / AGM region / differentiation potency / 胚幹細胞 / AGM / 分化能 / 発生運命 / ES細胞 / ImmortoMouse / 細胞の分化 / 細胞増殖と分化制御 / トランスジェニックマウス |
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| Research Abstract |
We have tried to establish hematopoietic stem cell lines by using a transgenic mouse strain (Immorto Mouse, Charles River Laboratory) harboring SV40 tsA 58 gene. To obtain the most primitive hamatopoietic stem cell population, we dissected out the yolk sac region from embryos at 8 days post coitum (dpc). Such isolated cells were cultured at 33ﾟC in the presence of interferon-gamma to promote transgene expression. They were maintained by subculturing every week for more than 6 months, during which cell cloning was performed. Such established cell lines were tested for their differentiation potency. So far, we obseved appearance of the macrophages and megakaryocytes.
As the second source of the early hematopoietic stem cells, we dissected out the ventral trunk region including the paraaortic splanchnopleura from embryos at 10.0 dpc. Recent reports indicate that this region produces the definitive hematopoietic stem cells. Microexplant culture of these tissues produced a cell line that has distinct cell morphology. In subculturing, these cells first produced an epithelia-like cell sheet, but at later stages, they produced hematoocyte-like small round cell population growing on top of the cell sheet. We are currently testing their differentiation potency.
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