1995 Fiscal Year Final Research Report Summary
Molecular study of hematopoiesis-supporting ability of C3H10T1/2 mouse embryo fibroblasts
| Project/Area Number |
06454345
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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 |
Hematology
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| Research Institution | Jichi Medical School |
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Principal Investigator |
OZAWA Keiya Jichi Medical School, Department of Medicine, Professor, 医学部, 教授 (30137707)
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| Co-Investigator(Kenkyū-buntansha) |
HONDA Hiroaki Jichi Med.Sch., Depart.of Medicine, Research Associate, 医学部, 助手 (40245064)
SAKAI Ryuichi Jichi Med.Sch., Depart.of Medicine, Research Associate, 医学部, 助手 (40215603)
MANO Hiroyuki Jichi Med.Sch., Depart.of Medicine, Associate Professor, 医学部, 助教授 (90240704)
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| Project Period (FY) |
1994 – 1995
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| Keywords | Stromal cell / Preadipocyte / C3H10T1 / 2 cell / Differentiation induction / Hematopoiesis-supporting ability / CD34 |
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| Research Abstract |
To investigate the molecular aspects of hematopoiesis-supporting ability of stromal cells, we used a differentiation-inducible mouse embryo fibroblast cell line, C3H10T1/2 (10T1/2). Stably determined preadipocyte and myoblast cell lines were established after a brief exposure of 10T1/2 cells to 5-azacytidine. These cell lines terminally differentiated into adipocytes and myotubes, respectively, under appropriate conditions. The hematopoiesis-supporting ability was significantly elevated at the preadipocyte stage (A54 preadipocyte), and was reduced after terminal adipocytic differentiation. To identify molecules that contribute to the hematopoiesis-supporting ability of preadipocytes, we screened genes that were differentially expressed in A54 preadipocytes and isolated the novel gene by mRNA differential display method. This gene was defined as a gene that was down-regulated during adipocyte differentiation-1 (drad-1). The drad-1 was expressed in other mouse preadipocytes, namely, ST2 and PA6 cells, that also have hematopoiesis-supporting ability. Moreover, the drad-1 was found to be expressed in mouse bone marrow. However, the function of the protein encoded by drad-1 is currently unknown. In addition, we investigated the change in CD34 mRNA expression during stromal cell differentiation. As a result, CD34 mRNA was constitutively expressed by parent 10T1/2 cells but not by adipogenically or myogenically determined cells. This finding supports the concept that CD34 may also be a marker of stromal progenitors and is lost as the cells differentiate into phenotypically distinct stromal elements. The 10T1/2-derived cell lines could provide a valuable tool to aid in the analysis of stromal cell development and differentiation and the search for novel stromal cell-derived factor (s).
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[Publications] Tsuruta, T., Tani, K., Shimane, M., Ozawa, K., Takahashi, S., Tsuchimoto, D., Takahashi, K., Nagata, S., Sato, N., Asano, S.: "Effects of myeloid cell growth factors on alkaline phosphatase, myeloperoxidase, defensin and granulocyte colony-stimulating factor receptor mRNA expression in hematopoietic cells of normal individuals and myeloid disorders." Brit.J.Haematol.(in press). (1996)
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[Publications] Suzuki, H., Suzuki, C., Konishi, A., Okano, A., Akiyama, Y., Ishibashi, T., Ozawa, K., and Asano, S.: "A synergistic increase in transplantable peripheral blood stem cells in mice by co-administration of recombinant human interleukin 6 and recombinant human granulocyte colony-stimulating factor." Transplantation. 59. 1596-1600 (1995)
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[Publications] Watari, K., Ozawa, K., Tajika, K., Tojo, A., Tani, K., Kamachi, S., Harigaya, K., Takahashi, T., Sekiguchi, S., Nagata, S., and Asano, S.: "Production of human granulocyte colony stimulating factor by various kinds of stromal cells in vitro detected by enzyme immunoassay and in situ hybridization." Stem Cells. 12. 416-423 (1994)
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[Publications] Yoshikubo, T., Ozawa, K., Takahashi, K., Nishikawa, M., Horiuchi, N., Tojo, A., Tani, K., Kodama, H., and Asano, S.: "Adhesion of NFS-60 myeloid leukemia cells to MC3T3-G2/PA6 stromal cells induces granulocyte colony-stimulating factor production." Blood. 84. 415-420 (1994)
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[Publications] Shimane, M., Tani, K., Maruyama, K., Takahashi, S., Ozawa, K., and Asano, S.: "Molecular cloning and characterization of G-CSF induced gene cDNA." Biochem.Biophys.Res.Commun.199. 26-32 (1994)
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