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Reprogramming of Human Fibroblasts to Induced Pluripotent Stem Cells with Sleeping Beauty Transposon-Based Stable Gene Delivery

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Transposons and Retrotransposons

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1400))

Abstract

Human induced pluripotent stem (iPS) cells are a source of patient-specific pluripotent stem cells and resemble human embryonic stem (ES) cells in gene expression profiles, morphology, pluripotency, and in vitro differentiation potential. iPS cells are applied in disease modeling, drug screenings, toxicology screenings, and autologous cell therapy. In this protocol, we describe how to derive human iPS cells from fibroblasts by Sleeping Beauty (SB) transposon-mediated gene transfer of reprogramming factors. First, the components of the non-viral Sleeping Beauty transposon system, namely a transposon vector encoding reprogramming transcription factors and a helper plasmid expressing the SB transposase, are electroporated into human fibroblasts. The reprogramming cassette undergoes transposition from the transfected plasmids into the fibroblast genome, thereby resulting in stable delivery of the reprogramming factors. Reprogramming by using this protocol takes ~4 weeks, after which the iPS cells are isolated and clonally propagated.

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Authors and Affiliations

  1. Division of Medical Biotechnology, Paul Ehrlich Institute, Paul Ehrlich Str. 51-59, 63225, Langen, Germany

    Attila Sebe & Zoltán Ivics Ph.D.

Authors
  1. Attila Sebe
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  2. Zoltán Ivics Ph.D.
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Correspondence to Zoltán Ivics Ph.D. .

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Editors and Affiliations

  1. Institute of Genetics and Molecular Medi, Genyo (Center for Genomics and Oncologic, Granada, Spain

    Jose L. Garcia-Pérez

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Sebe, A., Ivics, Z. (2016). Reprogramming of Human Fibroblasts to Induced Pluripotent Stem Cells with Sleeping Beauty Transposon-Based Stable Gene Delivery. In: Garcia-Pérez, J. (eds) Transposons and Retrotransposons. Methods in Molecular Biology, vol 1400. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3372-3_26

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  • DOI: https://doi.org/10.1007/978-1-4939-3372-3_26

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3370-9

  • Online ISBN: 978-1-4939-3372-3

  • eBook Packages: Springer Protocols

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