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Generation of induced pluripotent stem cells from a small amount of human peripheral blood using a combination of activated T cells and Sendai virus

Nature Protocols volume 7, pages 718–728 (2012)Cite this article

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Abstract

Induced pluripotent stem cells (iPSCs) have become important cell sources for genetic disease models, and they have the potential to be cell sources for future clinical therapies. However, invasive tissue sampling reduces the number of candidates who consent to donate cells for iPSC generation. In addition, integrated transgenes can potentially insert at inappropriate points in the genome, and in turn have a direct oncogenic effect. Technical modifications using a combination of activated T cells and a temperature-sensitive mutant of Sendai virus (SeV) can avoid invasive tissue sampling and residual transgene issues in generating iPSCs. Such advances may increase the number of consenting patients for cell donations. Here we present a detailed protocol for the generation of iPSCs from a small amount of human peripheral blood using a combination of activated T cells and mutant SeV encoding human OCT3/4, SOX2, KLF4 and c-MYC; T cell–derived iPSCs can be generated within 1 month of blood sampling.

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Figure 1: Overview of the TiPSC generation protocol.
Figure 2: Isolation and activation of PBMCs.
Figure 3: Infection of SeV.
Figure 4: Characterization of TiPSCs.

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Acknowledgements

T.S. is a research fellow of the Japan Society for the Promotion of Science. This work was supported in part by research grants from the Ministry of Education, Science and Culture, Japan, and by a grant from the New Energy and Industrial Technology Development Organization.

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Authors and Affiliations

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

    Tomohisa Seki, Shinsuke Yuasa & Keiichi Fukuda

  2. Department of Cardiology, Center for Integrated Medical Research, Keio University School of Medicine, Tokyo, Japan

    Shinsuke Yuasa

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  1. Tomohisa Seki
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  2. Shinsuke Yuasa
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  3. Keiichi Fukuda
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Contributions

T.S. and S.Y. prepared most of the paper. K.F. provided advice and proofread the paper.

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Correspondence to Shinsuke Yuasa or Keiichi Fukuda.

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The authors declare no competing financial interests.

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Seki, T., Yuasa, S. & Fukuda, K. Generation of induced pluripotent stem cells from a small amount of human peripheral blood using a combination of activated T cells and Sendai virus. Nat Protoc 7, 718–728 (2012). https://doi.org/10.1038/nprot.2012.015

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