| Project/Area Number |
10670303
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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 |
Immunology
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| Research Institution | Tottori University |
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Principal Investigator |
HAYASHI Shin-ichi Tottori University, Faculty of Medicine Professor, 医学部, 教授 (50208617)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1998: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | osteoclast / embryonic stem cell / hematopoiesis / M-CSF / ODF / OPGL / endothelial cell / stromal cell / macrophage / ES細胞 / tal-1 / GATA-2 |
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| Research Abstract |
Osteoclasts are hematopoietic cells essential for bone resorption. To understand whole process of osteoclastogenesis, I have developed a culture system with a stromal cell line, in which promotes differentiation of hematopoietic cell lineage, and terminal differentiation of osteoclasts from single embryonic stem (ES) cells.
This culture dispended with any passage or manipulation enabled us to investigate temporal and spatial localization of osteoclast lineage in the colonies from undifferentiated ES cells. On 4th day of cultures, hematopoietic cell and endothelial cell lineages arose in the central site of colonies. Cells expressed tartrate-resistant acid phosphatase (TRAPィイD1+ィエD1) which is a specific marker of osteoclast lineage were first detected on day 8 and subsequently localize at the edge of colonies and maturate into TRAPィイD1+ィエD1 multinucleated cells which have a potential of bone resorption.
A potential of osteoclastogenesis of gene targeted ES cell lines in tal-1, Gata-1, Gata-2 and Fog genes was examined. Gata-1(-/-) and Fog(-/-) ES cells differentiate into osteoclasts normally. No osteoclasts were detected in the culture of tal-1(-/-) ES cells. Gata-2(-/-) ES cells developed mature osteoclasts but the frequency reduced to one-tenth of normal ES cell lines. I also decided the ordered requirement for osteoclastogenesis as tal-1 --> Gata --> macrophage colony-stimulating factor --> osteoclast differentiation factor.
Recently, I established a new culture system for melanocyte development from ES cells. These systems are useful tools for study of osteoclastogenesis, because melanocytes are good internal control of cell dicision and culture conditions.
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