Simple Transportation of Genetically Engineered Mice via Cold Storage Techniques

  • Hidetaka Yoshimoto
  • Toru TakeoEmail author
  • Naomi Nakagata
Part of the Methods in Molecular Biology book series (MIMB, volume 2066)


Live genetically modified (GM) mice are often shipped in transportation cages. However, the shipment of live mice is sometimes associated with difficulties. Recently, we developed an alternative means to transport GM mice via cold storage techniques for sperm and embryos. Cold storage of the cauda epididymis, the male reproductive organ for storing mature sperm, can maintain the fertility of sperm in cold preservation medium for 10 days. The sperm can then be used to produce embryos or pups via in vitro fertilization and embryo transfer. Conversely, cold-stored two-cell embryos maintain developmental ability for 4 days. The embryos can be used to produce pups via embryo transfer. Cold transport techniques are being increasingly employed to create local and global networks between research institutes for the shipment of GM mice. In this text, we described a cold transport technique for sperm and two-cell embryos in mice.

Key words

Transportation Cold storage Cauda epididymis Two-cell embryo 



The authors thank Yuka Horikoshi, Shiori Takeuji, Kiyoko Yamashita, Tomoko Kondo, Yukie Haruguchi, Yumi Takeshita, Yuko Nakamuta, and Shuuji Tsuchiyama (CARD, Kumamoto University) for technical support. This work was partially supported by grants of the National Bioresource Project from the Ministry of Education, Culture, Sports, Science and Technology of Japan and Research on Development of New Drugs from the Japan Agency for Medical Research and Development.


  1. 1.
    Takeo T, Tsutsumi A, Omaru T, Fukumoto K, Haruguchi Y, Kondo T, Nakamuta Y, Takeshita Y, Matsunaga H, Tsuchiyama S, Sakoh K, Nakao S, Yoshimoto H, Shimizu N, Nakagata N (2012) Establishment of a transport system for mouse epididymal sperm at refrigerated temperatures. Cryobiology 65(3):163–168. Scholar
  2. 2.
    Yoshimoto H, Takeo T, Irie T, Nakagata N (2017) Fertility of cold-stored mouse sperm is recovered by promoting acrosome reaction and hyperactivation after cholesterol efflux by methyl-beta-cyclodextrin. Biology of Reproduction 96(2):446-455CrossRefGoogle Scholar
  3. 3.
    Yoshimoto H, Takeo T, Nakagata N (2017) Dimethyl sulfoxide and quercetin prolong the survival, motility, and fertility of cold-stored mouse sperm for 10 days†. Biology of Reproduction 97(6):883-891CrossRefGoogle Scholar
  4. 4.
    Takeo T, Hoshii T, Kondo Y, Toyodome H, Arima H, Yamamura K, Irie T, Nakagata N (2008) Methyl-beta-cyclodextrin improves fertilizing ability of C57BL/6 mouse sperm after freezing and thawing by facilitating cholesterol efflux from the cells. Biol Reprod 78(3):546–551. Scholar
  5. 5.
    Takeo T, Nakagata N (2011) Reduced glutathione enhances fertility of frozen/thawed C57BL/6 mouse sperm after exposure to methyl-beta-cyclodextrin. Biol Reprod 85(5):1066–1072. Scholar
  6. 6.
    Nakagata N, Takeo T, Fukumoto K, Kondo T, Haruguchi Y, Takeshita Y, Nakamuta Y, Matsunaga H, Tsuchiyama S, Ishizuka Y, Araki K (2013) Applications of cryopreserved unfertilized mouse oocytes for in vitro fertilization. Cryobiology 67(2):188–192. Scholar
  7. 7.
    Takeo T, Fukumoto K, Kondo T, Haruguchi Y, Takeshita Y, Nakamuta Y, Tsuchiyama S, Yoshimoto H, Shimizu N, Li MW, Kinchen K, Vallelunga J, Lloyd KC, Nakagata N (2014) Investigations of motility and fertilization potential in thawed cryopreserved mouse sperm from cold-stored epididymides. Cryobiology 68(1):12–17. Scholar
  8. 8.
    Takeo T, Nakagata N (2010) Combination medium of cryoprotective agents containing L-glutamine and methyl-{beta}-cyclodextrin in a preincubation medium yields a high fertilization rate for cryopreserved C57BL/6J mouse sperm. Lab Anim 44(2):132–137. Scholar
  9. 9.
    Takeo T, Kondo T, Haruguchi Y, Fukumoto K, Nakagawa Y, Takeshita Y, Nakamuta Y, Tsuchiyama S, Shimizu N, Hasegawa T, Goto M, Miyachi H, Anzai M, Fujikawa R, Nomaru K, Kaneko T, Itagaki Y, Nakagata N (2010) Short-term storage and transport at cold temperatures of 2-cell mouse embryos produced by cryopreserved sperm. J Am Assoc Lab Anim Sci 49(4):415–419PubMedPubMedCentralGoogle Scholar
  10. 10.
    Horikoshi Y, Takeo T, Nakagata N (2016) N-acetyl cysteine prolonged the developmental ability of mouse two-cell embryos against oxidative stress at refrigerated temperatures. Cryobiology 72(3):198–204. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Hidetaka Yoshimoto
    • 1
  • Toru Takeo
    • 1
    Email author
  • Naomi Nakagata
    • 1
  1. 1.Division of Reproductive Engineering, Center for Animal Resources and Development (CARD)Kumamoto UniversityChuo-ku, Kumamoti-shiJapan

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