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Derivation of iPSCs in stirred suspension bioreactors

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Abstract

Induced pluripotent stem cells (iPSCs) are typically derived in adherent culture. Here we report fast and efficient derivation of mouse iPSCs in stirred suspension bioreactors, with and without the use of c-Myc. Suspension-reprogrammed cells expressed pluripotency markers, showed multilineage differentiation in vitro and in vivo, and contributed to the germline in chimeric mice. Suspension reprogramming has the potential to accelerate and standardize iPSC research.

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Figure 1: Expansion and characterization of SiPSCs.
Figure 2: Developmental potential of 3F-SiPSCs.

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Acknowledgements

We acknowledge members of The Clara Christie Centre for Mouse Genomics for helping us to derive 129/Sv MEFs and SiPSC chimeras. This research was supported by a Canadian Institute of Health Research Regenerative Medicine and Nanomedicine Team grant (RMF-82497).

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Author notes

  1. Roman Krawetz and Derrick E Rancourt: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada

    Mehdi Shafa, Brad Day, Akihiro Yamashita, Guoliang Meng, Shiying Liu & Derrick E Rancourt

  2. Department of Surgery, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada

    Roman Krawetz

Authors
  1. Mehdi Shafa
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  2. Brad Day
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  3. Akihiro Yamashita
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  4. Guoliang Meng
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  5. Shiying Liu
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  6. Roman Krawetz
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  7. Derrick E Rancourt
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Contributions

M.S. designed the project, generated and characterized SiPSCs and prepared the manuscript; B.D. performed confocal microscopy and qPCR and prepared the manuscript; A.Y., G.M. and S.L. characterized SiPSCs, R.K. and D.E.R. designed the project and prepared the manuscript.

Corresponding author

Correspondence to Derrick E Rancourt.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–3, Supplementary Tables 1–6 (PDF 339 kb)

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Shafa, M., Day, B., Yamashita, A. et al. Derivation of iPSCs in stirred suspension bioreactors. Nat Methods 9, 465–466 (2012). https://doi.org/10.1038/nmeth.1973

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  • DOI: https://doi.org/10.1038/nmeth.1973

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