| Project/Area Number |
01440089
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory animal science
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| Research Institution | CENTRAL INSTITUTE FOR EXPERIMENTAL ANIMALS (CIEA) |
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Principal Investigator |
NOMURA Tatsuji CIEA, Director, 所長 (10072399)
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| Co-Investigator(Kenkyū-buntansha) |
TOYODA Yutaka Univ Tokyo, Inst Med Sci, Dept Reprod & Dev Biol, Professor, 獣医学研究部, 教授 (90050418)
ITO Mamoru CIEA, Immunol Lab, Chief Scientist, 免疫研究室, 室長 (00176364)
HASEGAWA Takanori CIEA, Dev Biotech Lab, Scientist, 発生工学研究室, 研究員 (20192271)
YOKOYAMA Minesuke CIEA, Reprod Lab, Chief Scientist, 生殖研究室, 室長 (40090930)
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| Project Period (FY) |
1989 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥29,900,000 (Direct Cost: ¥29,900,000)
Fiscal Year 1991: ¥7,300,000 (Direct Cost: ¥7,300,000)
Fiscal Year 1990: ¥8,000,000 (Direct Cost: ¥8,000,000)
Fiscal Year 1989: ¥14,600,000 (Direct Cost: ¥14,600,000)
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| Keywords | Laboratory Animals / Mice / Rats / Animal Models for Human Diseases / Embryos / Embryo Cryopreservation / Embryo Bank / Preservation of Strains / バイオサイエンス |
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| Research Abstract |
The goals of this research were the development and improvement of cryopreservation techniques for early embryos and gametes (spermatozoa) of mice and rats, and to establish an embryo bank system using these cryopreservation techniques. The results of this research are outlined below.
1) A cryopreservation method for mouse spermatozoa was developed and live young originating from cryopreserved spermatozoa were obtained successfully by in vitro fertilization. This report (Experimental Animals 39: 125-128, 1990) was the first successful case in the world using mouse spermatozoa.
2) The numbers of cases of spontaneous ovulation and induced ovulation by hormone (PNSG-hCG) treatment in various inbred strains, hybrid strains and closed colonies were investigated and conditions for the efficient collection of embryos from these strains were determined.
3) Fertilized eggs were prepared by in vitro fertilization and a system for obtaining embryos for cryopresevation systematically was established.
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With this system, it became possible to obtain fertilized eggs and embryos from strains of animal models with reproductive disorders.
4) By means of a system of cryopreservation of mouse embryos combining hormone treatment and in vitro fertilization, 2-cell stage embryos of more than 70 strains maintained in CIEA or introduced from research institutes in Japan and overseas, as well as about 500 strains of transgenic mice created in CIEA, were cryopreserved. As of the end of March 1992, about 170,000 mouse embryos had been cryopreserved.
5) Cryopreservation methods for 2-cell stage embryos of rats were studied and techniques which can be applied in cryopreservation have been established. These are basically improvements of the methods used for mice, and the recovery rate of frozen and thawed embryos to live young is about 50-60%, the same as that for mice.
6) Two-cell stage embryos of about 40 strains of rats maintained in CIEA or introduced from institutes in Japan and overseas have been cryopreserved. As of the end of March 1992, about 5,000 rat embryos have been cryopreserved.
7) A system which can serve as an embryo bank consisting of liquid nitrogen storage containers for preservation of large cryopreservation samples has been established. With this system, it is possible to preserve more than 500,000 embryos in 36,000 tubes. Based on these results, a system which can serve as an embryo bank for mice and rats has been established in CIEA. In the future, it will be necessary to study the software required for the functional operation of this embryo bank. Less
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