Brief Communication

A nonviral minicircle vector for deriving human iPS cells

  • Nature Methods volume 7, pages 197199 (2010)
  • doi:10.1038/nmeth.1426
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Abstract

Owing to the risk of insertional mutagenesis, viral transduction has been increasingly replaced by nonviral methods to generate induced pluripotent stem cells (iPSCs). We report the use of 'minicircle' DNA, a vector type that is free of bacterial DNA and capable of high expression in cells, for this purpose. Here we use a single minicircle vector to generate transgene-free iPSCs from adult human adipose stem cells.

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Acknowledgements

We thank A.J. Connolly for assistance with histological analysis, members of the Stanford Functional Genomics Facility and Stanford University PAN Core Facility for assistance with microarrays and A. Cherry for assistance with cytogenetics. We thank funding support from Mallinckrodt Foundation, US National Institutes of Health (NIH) DP2OD004437, HL091453-01A1S109, Burroughs Wellcome Foundation and American Heart Association 0970394N (J.C.W.); NIH R90 DK 07010301, California Institute of Regenerative Medicine T1-00001 and RL1-00662-1, NIH R21 DE018727, RC1HL100490, NIH R21 DE019274, the Oak Foundation and the Hagey Laboratory for Pediatric Regenerative Medicine (M.T.L.); U01HL099776 (R.C.R.).

Author information

Affiliations

  1. Departments of Medicine and Radiology, Stanford University School of Medicine, Stanford, California, USA.

    • Fangjun Jia
    • , Kitchener D Wilson
    • , Ning Sun
    • , Mei Huang
    • , Zongjin Li
    •  & Joseph C Wu
  2. Department of Bioengineering, Stanford University School of Medicine, Stanford, California, USA.

    • Kitchener D Wilson
  3. Department of Surgery, Stanford University School of Medicine, Stanford, California, USA.

    • Deepak M Gupta
    • , Nicholas J Panetta
    •  & Michael T Longaker
  4. Departments of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, California, USA.

    • Zhi Ying Chen
    •  & Mark A Kay
  5. Department of Cardiothoracic Surgery, Stanford University School of Medicine, Stanford, California, USA.

    • Robert C Robbins
  6. Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California, USA.

    • Michael T Longaker
    •  & Joseph C Wu

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Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Mark A Kay or Michael T Longaker or Joseph C Wu.

Supplementary information

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    Supplementary Text and Figures

    Supplementary Figures 1–9 and Supplementary Tables 1–4

Videos

  1. 1.

    Supplementary Video 1

    Day 20 beating cardiomyocyte progenitors derived from mc-iPSCs. We initially observed beating clusters on day 16 after EB formation. Two clusters of cells can be seen beating spontaneously in this video (lower right corner and middle top).