| Project/Area Number |
11680818
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Laboratory animal science
|
|---|
| Research Institution | The University of Tokyo |
|---|
Principal Investigator |
SUDO Katsuko Institute of Medical Science, Center for Experimental Medicine, The University of Tokyo Research associate, 医科学研究所, 助手 (50126091)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
SHIBATA Shinnwa Research fellow of Japan Society for the Promotion of Scientist, 特別研究員(PD)
ASANO Masahide Institute for Experimental animals, Graduate school of Medical Science, Kanazawa University. Professor, 医学部, 教授 (50251450)
IWAKURA Yoichiro Institute of Medical Science, Center for Experimental Medicine, The University of Tokyo Professor, 医科学研究所, 教授 (10089120)
|
|---|
| Project Period (FY) |
1999 – 2000
|
| Project Status |
Completed (Fiscal Year 2001)
|
| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2000: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1999: ¥1,900,000 (Direct Cost: ¥1,900,000)
|
|---|
| Keywords | transgenic mice / SDF-1 / stem cell / scid mice |
|---|
| Research Abstract |
Mice with severe combined immunodeficiency (scid) mutation support the development of human hematopoietic stem cells to all lineages of blood cells except for T-lymphocytes. Several lines of evidences suggest that the failure of human blood stem cells to develop o T-cells in mice can be explained by their incapable homing to the thymus of mice. In order to generate recipient mice which support the development of human hematopoietic stem cells to T-cells, we intended to generate transgenic mice that express SDF-1, a chemoattractant factor of blood stem cells, in the thymus.
We adopted the DNA microinjection technology into fertilized eggs to obtain the transgenic mice. We prepared two DNA constructs ; one consisted of humanized SDF-1 gene under control of proximal lck promoter and human growth hormine poly-A genomic region, and other of human FasL ene with a point mutation under control of the same promoter and poly-A region. The mutant FasL. which transmits death signal only to cell of
… More
mice but not to human cells, was utilized to avoid the development of thymic lymphoma in scid mice. These two DNA constructs were injected at the same time. We injected the DNA constructs into 699 fertilized eggs of C3H/HeN X NOD-scid or NOD-scid mice, and 74pups were obtained. Among them, 9 mice were transgenic mice and one of them successfully expressed the transgene. When analyzed the F1 transgenic mouse of scid/scid genotype by northern blotting, it Expressed the transgene in the thymus and spleen despite it was not received irradiation. Since thymocytes of scid mice halt their development in CD4-CD8-stage and genes under the control of lck promoter are activated after CD4+CD8+stage, the expression of transgenes in the thymus of scid mice is suggested to be due to the leakey phenotype. We have successfully generated a line of transgenic mice that express humanized SDF-1, a chemoattractant of blood tem cells, in the thymus. This mouse is expected to support the development of human hematopoietic stem cells to T-cells when transferred, and may be useful animal model for studying human hematopoiesis, immunology, and leukemia. Less
|
