| Project/Area Number |
07558290
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Developmental biology
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| Research Institution | National Institute of Genetics |
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Principal Investigator |
NAKATSUJI Norio National Institute of Genetics, Mammalian Development Laboratory, Professor, 系統生物研究センター, 教授 (80237312)
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| Co-Investigator(Kenkyū-buntansha) |
OGURA Atsuo National Institute of Infectious Diseases, Department of Veterinary Science, Sen, 獣医科学部, 主任研究官 (20194524)
SHIRAYOSHI Yasuaki National Institute of Genetics, Mammalian Development Laboratory, Assistant Prof, 医学部, 助教授 (90249946)
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| Project Period (FY) |
1995 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1997: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1996: ¥3,400,000 (Direct Cost: ¥3,400,000)
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| Keywords | mouse / primordial germ cells / gene transfection / apoptosis / EG cells / artificial fertilization / spermatocyte / transgenic mouse / 生殖細胞 / 性分化 / 減数分裂 / 生殖巣 / 胚移植 / 胎仔生殖細胞 / 体外受精 / 精子細胞 / 顕微注入 / 細胞融合 / 細胞増殖 |
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| Research Abstract |
To develop novel methods of reproductive and genetical manipulation of mammalian fetal germ cells, we carried out the in vitro culture, gene transfection, and micromanipulation of mouse fetal germ cells. Firstly, we improved culture conditions of primordial germ cells. We then tested various methods of gene transfection into cultured fetal germ cells. We observed inhibition of apoptosis by transfection of bcl-x gene and adenovirus E1b 19k gene. We also tested differentiation capacity of EG cells established by addition of forskolin and retinoic acid in culture of primordial germ cells. After transplantation into the W mutant mouse testis, they showed differentiation capacity of ES or teratoma cells but not the germ cells.
Also, new micromanipulation methods were developed to produce offspring from germ cells in various artificial fertilization procedures. We first improved the method of using round spermatid for production of fertilized eggs by micromanipulation. We also succeeded to produce normal diploid embryos by using primary spermatocytes and ovum. During the microsurgical fertilization using spermatid, a GFP gene solution was injected at the same time. Such developed blastocytes showed expression of the GFP fluorescence, thus indicating possibility of the gene transfection using such procedure.
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