| Project/Area Number |
04557034
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
内科学一般
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| Research Institution | University of Tsukuba (1994)
The Institute of Physical and Chemical Research (1992-1993) |
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Principal Investigator |
NAKAUCHI Hiromitsu (1992, 1994) Univ.of Tsukuba Dept.of Immunology, Professor, 基礎医学系, 教授 (40175485)
中村 幸夫 (1993) 理化学研究所, 造血制御研究チーム, 研究員 (60231479)
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| Co-Investigator(Kenkyū-buntansha) |
TANI Kenzaburo Institute of Medical Science Univ.of Tokyo, Assitant Professor, 医科学研究所, 講師 (00183864)
HIROCHIKA Rei RIKEN Lab.of Cell Growth and Differentiation, Staff Scientist, 造血制御研究チーム, 研究員 (90260223)
NAKAMURA Yukiko Univ.of Tsukuba Dept.of Immunology, Assitant Professor, 基礎医学系, 講師 (60231479)
徳元 康人 筑波大学, 基礎医学系, 助手 (70261170)
中内 啓光 筑波大学, 基礎医学系・免疫学, 教授 (40175485)
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| Project Period (FY) |
1992 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥15,800,000 (Direct Cost: ¥15,800,000)
Fiscal Year 1994: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1993: ¥6,100,000 (Direct Cost: ¥6,100,000)
Fiscal Year 1992: ¥7,200,000 (Direct Cost: ¥7,200,000)
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| Keywords | somatic cell gene therapy / hematopoietic stem cells / cytokines / diabetes / bone marrow transplantation / CD34 / 多能性 / 自己複製能 / c-kit / 免疫寛容 / clonal deletion / clonal anergy / 骨髄キメラ / 遺伝子治療 / 血液幹細胞 / インシュリン / アガロース |
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| Research Abstract |
1)Establishment of a mouse model system of a somatic cell gene therapy with an immunological safety system.
Mouse fibroblasts were transfected with an expression vector containing human insulin cDNA.When one million of these fibroblasts were injected i.p.into streptozocin-induced diabetic mice, we observed a significant improvement of the blood glucose concentrations. To ensure the safety of somatic cell gene therapy, the proinsulin-producing fibroblasts were further transfected with an expression vector containing CD8.2, a gene encoding a cell surface antigen.Somatic gene therapy with these doubly transfected cells followed by the administration of a monoclonal antibody to CD8.2 resulted in an initial decrease of blood glucose concentrations followed by the permanent recurrence of hyperglycemia, thus proving the complete removal of the transplanted cells.
2)Purification of the hematopoietic stem cells in adult mouse bone marrow.
Using monoclonal antibodies and FACS,we established a system by which the hematopoietic stem cells can be isolated. The results indicate that c-kit+Sca-1+CD34-Lin-cells constituting only 0.008% of bone marrow mononuclear cells are highly enriched for long-term marrow repopulating ability. We were able to reconstitute bone marrow of a lethally irradiated mouse by an injection of a single c-kit+Sca-1+CD34-Lin-cell. We also demonstrated that short-term culture of enriched stem cells under certain cytokines increase the efficiency of retroviral transdution into the hematopoietic stem cells.
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