| Project/Area Number |
59480153
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Experimental pathology
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| Research Institution | Keio University |
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Principal Investigator |
HARIGAYA Kenichi (1985-1986) School of Medicine, Keio University, 医学部, 助手 (40101894)
張ヶ谷 健一 (1984) 慶応義塾大学, 医学部, 助手
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| Co-Investigator(Kenkyū-buntansha) |
HANDA Hiroshi The Institute of Medical Science, University of Tokyo, 医科学研究所, 助教授 (80107432)
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| Project Period (FY) |
1984 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1986: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1985: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1984: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Keywords | Bone marrow stromal cell line / Recombinant plasmid / Hematopoietic factor Hematopoietic stem cells / Cell to cell interaction / アミノ酸分析 / 細胞間相互作用 / 造血幹細胞 / 増殖因子 |
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| Research Abstract |
Five functional clonal cell lines were established from the adherent cell populations in long-term liquid cultures after transfection with the recombinant plasmid pSV3gpt. The radioimmunoassay and northern blot analysis revealed that these cell lines produced granulopoietic colony-stimulating factor (G-CSF) and macrophage colony-stimulating factor (M-CSF). The conditioned medium from the cell line KM-102 contained a factor which stimulated the proliferation and differentiation of multipotent stem cells. This factor was purified 340000 folds by five sequential fractionations and gave a single broad band of protein with a molecular weight of approximately 18000 dalton. This purified factor was associated with burst promoting activity (BPA) in vitro and showed is a highest specific activity of BPA among the substances which have been reported so far. The N-terminal amino acid sequence of this factor was analogous to the one deduced from cDNA of human GM-CSF. A part of human hemic cell lin
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es, HL-60 and KG-1, attached to the marrow stromal cell lines. Furthermore, the proliferation and differentiation of the attached hemic cells were strongly inhibited even when the cocultures were supplemented with differentiation inducing agent, 1 <alpha> , <25(OH)_2> vitamin <D_3> . These attachment is specific between myeloid cells and marrow stromal cells since there was no binding between myeloid cell lines and stromal cells from human thymus and lymph nodes, and between T or B lymphocytic cell lines and the marrow stromal cells. These indicate that direct cell to cell interaction is present between myeloid hemic cells and marrow stromal cells. Gap junctional communication was frequently observed in primary marrow stromal cells and stromal cell lines, and sometimes between stromal cells and the attached round hemic cells. This suggests that direct communication of low molecular weight substances is present between these cells. Nevertheless, the nature of the communication has not been clarified. Less
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