1991 Fiscal Year Final Research Report Summary
A Research for cryopreservation and transportation of embryos of small lsboratory animals.
| Project/Area Number |
02558008
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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 |
Laboratory animal science
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| Research Institution | Hokkaido University |
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Principal Investigator |
KASAI Noriyuki Hokkaido University School of Medicine, Associate Professor, 医学部, 助教授 (60001947)
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| Co-Investigator(Kenkyū-buntansha) |
YAMASHITA Tadashi Hokkaido University School of Veterinary Medicine, Instructor, 獣医学部, 助手 (30220338)
MIYOSHI Ichiro Hokkaido University School of Medicine, Instructor, 医学部, 助手 (10183972)
HIRABAYASHI Masumi Institute for biological science, Yukijirushi Milk Co. Ltd., Research Investigat, 生物科学研究所, 研究員
KASAI Magosaburo Kochi University Faculty of Agriculture, Associate Professor, 農学部, 助教授 (60152617)
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| Project Period (FY) |
1990 – 1991
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| Keywords | Mouse / Rat / Embryo transport / Embryo cryopreservation / Non-freezing technique / Vitrification method / Microbiological level / Forwarding agent |
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| Research Abstract |
We attempted to develop two embryo transport systems using nonfreezing techniques and using a combination of vitrification techniques and a non-liquid type liquid nitrogen container. For nonfreezing technique, mouse morulae were kept in 0.5M Sucrose-PB1 solution in a thermos bottle with ice-water, followed by packaging with polystyrene foam. For vitrification technique, mouse morulae were kept in 40% ethylene glycol, 30% Ficoll and 0.5M sucrose in a 0.25ml plastic straw, followd by immediate immersing in liquid nitrogen and packaging in a non-liquid type ultralow temperature container with a special shipping enclosure. The packages were transported from Kochi to Sapporo, about 1300km by a forwarding agent. All packages arrived at the destination in 48 hours. After shipping, the survival of embryos was 46.8% for nonfreezing technique and 88.0% for vitrification techniques when evaluated by development to blastocysts in vitro. A final birthrate was 14.1% and 45.8% for nonfreezing techniques and vitrification techniques, respectively. Both systems are cheap, convenient and practical for embryo transports, and can be selected by the situation of sending and receiving facilities.
We also attempted to improved the preservation technique by vitrification for mouse embryos and rat embryos. These improved preservation techniques are very simple and much helpful for prevailling embryo preservation and transport system.
We also showed embryo transfer technique can clear up of infections organisms from conventional animals, indicating that we may disregard the difference of microbiogical level between the donor and recipient laboratories owing to the intrinsic aseptic condition of treated embryos.
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[Publications] Kasai, M., Komi, J. H., Takakamo, A., Tsudera, H., Sakurai, T., and Machida, T.: "A simple method for mouse embryo cryopreservation in a low toxicity vitrification solution, without appreciable loss of viability." J. Reprod. Fert. 89. 91-97 (1990)
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[Publications] Kasai, M., Hamaguchi, Y., Zhu, S. E., Miyabe, T., Sakurai, T., and Machida, T.: "High survival of rabbit morulae after vitrification in an ethylene glycol-based solution by a simple method." Biology of Reproduction.(1992)
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[Publications] Miyoshi, I., Yamashita, T., Aoyama, S., Sasaki, N., Maki, K., Yamada, S., and Kasai, N.: "Effect of environmental factors on the yield of mouse embryos superovulated by hormone-induction." Lab. Anim.(1992)
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