| Project/Area Number |
18500336
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory animal science
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| Research Institution | Central Institute for Experimental Animals |
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Principal Investigator |
SASAKI Erika Central Institute for Experimental Animals, Marmoset Research Departmenet, Laboratory of Applied Developmental Biology, Lab Head (70390739)
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| Co-Investigator(Kenkyū-buntansha) |
SOTOMARU Yusuke Hiroshima University, Natural Science Center for Basic research and Deverlopment, Associate professor (90309352)
江藤 智生 財団法人実験動物中央研究所, 動物資源管理部生殖工学研究室, 室長代理 (30370175)
平川 玲子 財団法人実験動物中央研究所, マーチセット研究部, 共同研究員 (70414068)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥4,110,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥510,000)
Fiscal Year 2007: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2006: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | Nuclear transfer / Developmental Biology / Primates / 再生医学 / 発生・分化 / scntES細胞 / 体細胞核移植 |
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| Research Abstract |
The nuclear transfer technique has been applied to various mammals to produce clone animals, but a standardized method is not applicable to all species. We examined here the optimum procedure for somatic cell cloning in common marmosets. In a preliminary experiment, we confirmed that parthenogenetic activation of in vitro matured oocytes was successfully induced by electrical stimulation (3 cycles of 150 V/mm, 50 μsec x 2, 20min intervals), and this condition was applied to the following experimental procedure. Using marmoset embryonic fibroblast cells as donor nuclei, nuclear transfer to recipient enucleated oocytes was performed 1h before, immediately after or 1h after activation treatment. After in vitro culture for 10 days, development to blastocyst stage was observed in only oocytes of which nuclei were transferred 1h before activation (4.3%), and in no other cases. We then examined the influence of the derivation of donor cells on development to blastocyst stage. Clone embryos were produced from donor myeloid cells from male and female adults and skin-derived fibroblasts from male adults. As a result, development to blastocyst stage was observed in 9.4% (myeloid from female), 14.5% (myeloid from male) and 9.3% (fibroblast from female) of clone embryos. Somatic cell cloned embryo with developmental ability to blastocyst stage is constructed when the donor cells have been transferred into the recipient, prior to activation.
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