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Derivation of Induced Pluripotent Stem Cells by Retroviral Gene Transduction in Mammalian Species

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Genomic Imprinting

Part of the book series: Methods in Molecular Biology ((MIMB,volume 925))

Abstract

Pluripotent stem cells can provide us with an enormous cell source for in vitro model systems for development. In 2006, new methodology was designed to generate pluripotent stem cells directly from somatic cells, and these cells were named induced pluripotent stem cells (iPSCs). This method consists of technically simple procedures: donor cell preparation, gene transduction, and isolation of embryonic stem cell-like colonies. The iPSC technology enables cell biologists not only to obtain pluripotent stem cells easily but also to study the reprogramming events themselves. Here, we describe the protocols to generate iPSCs from somatic origins by using conventional viral vectors. Specifically, we state the usage of three mammalian species: mouse, common marmoset, and human. As mouse iPSC donors, fibroblasts are easily prepared, while mesenchymal stem cells are expected to give rise to highly reprogrammed iPSCs efficiently. Common marmoset (Callithrix jacchus), a nonhuman primate, represents an alternative model to the usual laboratory animals. Finally, patient-specific human iPSCs give us an opportunity to examine the pathology and mechanisms of dysregulated genomic imprinting. The iPSC technology will serve as a valuable method for studying genomic imprinting, and conversely, the insights from these studies will offer valuable criteria to assess the potential of iPSCs.

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Acknowledgements

This work was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT); the Ministry of Health, Labor, and Welfare; the Japan Society for the Promotion of Science (JSPS); the National Institute of Biomedical Innovation; the Project for Realization of Regenerative Medicine, MEXT; the Funding Program for World-leading Innovative R&D in Science and Technology (FIRST), JSPS; and Grant-in-Aid for Young Scientists (B).

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Authors and Affiliations

  1. Department of Physiology, School of Medicine, Keio University, Tokyo, Japan

    Masanori Imamura, Hironobu Okuno, Ikuo Tomioka, Yoshimi Kawamura, Zachary Yu-Ching Lin, Ryusuke Nakajima, Wado Akamatsu, Hirotaka James Okano, Yumi Matsuzaki, Erika Sasaki & Hideyuki Okano

  2. Laboratory of Applied Developmental Biology, Marmoset Research Department, Central Institute for Experimental Animals, Kawasaki, Japan

    Ikuo Tomioka & Erika Sasaki

  3. PRESTO Japan Science and Technology Agency, Tokyo, Japan

    Erika Sasaki

Authors
  1. Masanori Imamura
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  2. Hironobu Okuno
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  3. Ikuo Tomioka
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  4. Yoshimi Kawamura
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  5. Zachary Yu-Ching Lin
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  6. Ryusuke Nakajima
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  7. Wado Akamatsu
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  8. Hirotaka James Okano
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  9. Yumi Matsuzaki
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  10. Erika Sasaki
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  11. Hideyuki Okano
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Corresponding author

Correspondence to Hideyuki Okano .

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Editors and Affiliations

  1. School of Medicine, Fels Institute, Temple University, N. Broad Street 3307, Philadelphia, 19140, Pennsylvania, USA

    Nora Engel

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© 2012 Springer Science+Business Media, LLC

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Imamura, M. et al. (2012). Derivation of Induced Pluripotent Stem Cells by Retroviral Gene Transduction in Mammalian Species. In: Engel, N. (eds) Genomic Imprinting. Methods in Molecular Biology, vol 925. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-011-3_2

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  • DOI: https://doi.org/10.1007/978-1-62703-011-3_2

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-010-6

  • Online ISBN: 978-1-62703-011-3

  • eBook Packages: Springer Protocols

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