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Organ Generation from Knockedout Rat Blastocysts Complemented with Pluripotent Stem Cells

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Microinjection

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1874))

Abstract

Regeneration of human organs in domestic animal model would provide enough number of functional donor organs in transplantation therapy. Recent progresses in pluripotent stem cells and nuclease-based genome editing tools have set the stage for investigating the chimeric complementation approach to generate functional organs from embryonic stem (ES) cells or induced pluripotent stem (iPS) cells. In this chapter, protocol for allogeneic or xenogeneic organ generation using knocked-out (KO) rat blastocysts and the rat or mouse ES/iPS cells is described. The protocol includes (1) the preparation of KO rat colony, (2) the preparation of rat or mouse ES/iPS cells, (3) the recovery of rat blastocysts, (4) the stem cell injection into blastocysts, (5) the embryo transfer into pseudopregnant recipient uteri, and (6) the genotyping and organogenetic analysis of chimeric offspring. The accumulation of basic and practical knowledge in the rodent model would be useful in improving therapeutic performance to regenerate 3D organs available for transplantation.

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Author information

Authors and Affiliations

  1. The Graduate University for Advanced Studies, Okazaki, Aichi, Japan

    Masumi Hirabayashi

  2. Section of Mammalian Transgenesis, Center for Genetic Analysis of Behavior, National Institute for Physiological Sciences, Okazaki, Aichi, Japan

    Masumi Hirabayashi

  3. Faculty of Textile Science and Technology, Shinshu University, Ueda, Nagano, Japan

    Shinichi Hochi

Authors
  1. Masumi Hirabayashi
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  2. Shinichi Hochi
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Corresponding author

Correspondence to Masumi Hirabayashi .

Editor information

Editors and Affiliations

  1. National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA

    Chengyu Liu

  2. National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA

    Yubin Du

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Hirabayashi, M., Hochi, S. (2019). Organ Generation from Knockedout Rat Blastocysts Complemented with Pluripotent Stem Cells. In: Liu, C., Du, Y. (eds) Microinjection. Methods in Molecular Biology, vol 1874. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8831-0_18

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  • DOI: https://doi.org/10.1007/978-1-4939-8831-0_18

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8830-3

  • Online ISBN: 978-1-4939-8831-0

  • eBook Packages: Springer Protocols

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