| Project/Area Number |
03556038
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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 |
畜産学(含草地学)
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
MIYAMOTO Hazime Kyoto Univ.Fac.Agr.Prof., 農学部, 教授 (00026618)
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| Co-Investigator(Kenkyū-buntansha) |
KASAI M Kochi Univ.Fac.Agr.Prof., 農学部, 教授 (60152617)
NIWA K Okayama Univ.Fac.Agr.Prof., 農学部, 教授 (40089115)
SATO E Univ.Tokyo Inst.Med.Sci.Ass.Prof., 医科学研究所, 助教授 (80093243)
ISHII T Kyoto Univ.Fac.Agr.Instr., 農学部, 助手 (70111945)
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| Project Period (FY) |
1991 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥9,700,000 (Direct Cost: ¥9,700,000)
Fiscal Year 1993: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1992: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1991: ¥5,600,000 (Direct Cost: ¥5,600,000)
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| Keywords | Ovary / Bovine oocyte / Cryopreservation / Vitrification / Culture / In vitro fertilization / Ethylene glycol / Embryo transfer / 低温保存 / ヤギ卵巣 / ラット卵巣 / ウシ卵細胞 / マウス胚 / 緩慢凍結 / 還流培養 / 卵 / 卵胞破裂 / 凍結保存 |
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| Research Abstract |
1. A system has been developed for the parfusion of the goat ovary in vitro. The rupture of the Graafian follicle scarcely occurred. Glucose consumption and lactate production were observed as indices of cellular metabolic activity during perfusion.
2. Rat ovaries after vitrified in liquid nitrogen were transplanted to recipients. The growth of unilateral or bilateral ovaries were observed in 12 of 17 (71%) with grafts, and 6 of 12 (50%) rats displayd the regular estrous cycle. However, ovary cryopreservation of farm animals was impossible under the present conditions.
3. The highest survival rate was obtained when mouse embryos were vitrified at -196。C in the phosphate-buffered solution containing 40% ethylene glycol, 30% Ficoll and 0.5M sucrose. Mouse embryos vitrified developed into live young (61%). This simple and efficient method of cryopreservation will be applicable to embryos and oocytes of various species.
4. For development to the blastocyst stage of bovine oocytes fertilized in vitro, it is essential to include amino acids, phosphate, pyruvate and lactate in the medium, and glucose is necessary for later stages of development.
5. Bovine blastocysts were derived by fertilization of oocytes and development in vitro. Embryos were exposed to the vitrification solution at room temperature for 2 min and vitrified in liquid nitrogen. Then two embryos were transferred into each of ten recipient cows, and 8 (80%) became pregnant and 5 (25%) calves were born alive. In conclusion, the present system of in vitro fertilization of oocytes and cryopreservation by vitrification method is applicable to the effective utilization of oocytes in farm animals.
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