Cryobanking and Recovery of Genetically Modified Mice

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


Cryobanking of sperm, oocytes, and embryos is a useful means to efficiently maintain mouse colonies without breeding live animals. Cryopreserved cells can be permanently stored in well-managed systems in liquid nitrogen tanks at −196 °C and quickly reanimated for use via in vitro fertilization and/or embryo transfer. Recent improvements of reproductive technology markedly enhanced the efficiency of recovering and producing animals using cryopreserved cells. The establishment of a cryobanking system will increase the performance of animal experiments, meet the principles of 3Rs (replacement, reduction, and refinement), and reduce labour and costs. In this chapter, we described the latest techniques of sperm cryopreservation, in vitro fertilization, and oocyte and two-cell embryo vitrification developed at the Center for Animal Resources and Development (CARD).

Key words

Sperm cryopreservation In vitro fertilization Oocyte vitrification Two-cell embryo vitrification Embryo transfer 



We thank our staff, namely Kiyoko Yamashita, Tomoko Kondo, Yukie Haruguchi, Yumi Takeshita, Yuko Nakamuta, Tomoko Umeno, and Eri Ishida, and our students Hidetaka Yoshimoto and Ayumi Mukunoki 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.


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Copyright information

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

Authors and Affiliations

  1. 1.Division of Reproductive Engineering, Center for Animal Resources and Development (CARD)Kumamoto UniversityChuo-ku, Kumamoto-shiJapan

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