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Gene Transfer into Pluripotent Stem Cells via Lentiviral Transduction

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Embryonic Stem Cell Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1341))

Abstract

Recombinant lentiviral vectors are powerful tools to stably manipulate human pluripotent stem cells. They can be used to deliver ectopic genes, shRNAs, miRNAs, or any possible genetic DNA sequence into diving and nondividing cells. Here we describe a general protocol for the production of self-inactivating lentiviral vector particles and their purification to high titers by either ultracentrifugation or ultrafiltration. Next we provide a basic procedure to transduce human pluripotent stem cells and propagate clonal cell lines.

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Authors and Affiliations

  1. Institute of Clinical Biochemistry, Hannover Medical School, 30625, Hannover, Germany

    Ortwin Naujok, Ulf Diekmann & Matthias Elsner

Authors
  1. Ortwin Naujok
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  2. Ulf Diekmann
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  3. Matthias Elsner
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Corresponding author

Correspondence to Ortwin Naujok .

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Editors and Affiliations

  1. Ottawa Hospital Research Institute, Ottawa, Ontario, Canada

    Kursad Turksen

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© 2015 Springer Science+Business Media New York

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Naujok, O., Diekmann, U., Elsner, M. (2015). Gene Transfer into Pluripotent Stem Cells via Lentiviral Transduction. In: Turksen, K. (eds) Embryonic Stem Cell Protocols. Methods in Molecular Biology, vol 1341. Humana Press, New York, NY. https://doi.org/10.1007/7651_2015_221

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  • DOI: https://doi.org/10.1007/7651_2015_221

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2953-5

  • Online ISBN: 978-1-4939-2954-2

  • eBook Packages: Springer Protocols

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