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Protocols for Cryopreservation and Rederivation of Rat Gametes

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Rat Genomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2018))

Abstract

New genome-editing tools, such as ZFNs, TALEN, and CRISPR/Cas9, have enabled the generation of gene-modified models effectively in mammals. These technologies are a powerful tool for studying gene function and creating animal models for human diseases. On the other hand, such gene-modified animals are raised in numerous experimental animal facilities, which puts pressure on breeding space and maintenance costs. Embryo and sperm cryopreservation is not only the most simple and cost-effective method available for most gene-modified strains but also the most reliable method to preserve strains to avoid breeding problems and contamination. We have established a reliable, high quality embryo and sperm cryopreservation system for rat strains, ensuring the longevity of these valuable resources for the scientific community. These cryopreserved resources have been successfully used to rederive next generation pups using embryo transfer and intracytoplasmic sperm injection (ICSI). In this chapter, we describe in detail protocols for rat embryo vitrification and sperm cryopreservation followed by pup rederivation using the ICSI procedure and embryo transfer.

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Acknowledgments

This work was supported by the National Heart, Lung, and Blood Institute of the National Institutes of Health (R24HL114474).

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Authors and Affiliations

  1. Department of Physiology, Genomic Sciences and Precision Medicine Center, Medical College of Wisconsin, Milwaukee, WI, USA

    Akiko Takizawa

  2. Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan

    Tomoo Eto

Authors
  1. Akiko Takizawa
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  2. Tomoo Eto
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Corresponding author

Correspondence to Akiko Takizawa .

Editor information

Editors and Affiliations

  1. Department of Biomedical Engineering, Rat Genome Database, Medical College of Wisconsin, Milwaukee, WI, USA

    G. Thomas Hayman

  2. Department of Biomedical Engineering, Rat Genome Database, Medical College of Wisconsin, Milwaukee, WI, USA

    Jennifer R. Smith

  3. Department of Physiology, Rat Genome Database, Genomic Sciences and Precision Medicine Center, Medical College of Wisconsin, Milwaukee, WI, USA

    Melinda R. Dwinell

  4. Department of Biomedical Engineering, Rat Genome Database, Medical College of Wisconsin, Milwaukee, WI, USA

    Mary Shimoyama

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Takizawa, A., Eto, T. (2019). Protocols for Cryopreservation and Rederivation of Rat Gametes. In: Hayman, G., Smith, J., Dwinell, M., Shimoyama, M. (eds) Rat Genomics. Methods in Molecular Biology, vol 2018. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9581-3_6

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  • DOI: https://doi.org/10.1007/978-1-4939-9581-3_6

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9580-6

  • Online ISBN: 978-1-4939-9581-3

  • eBook Packages: Springer Protocols

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