A nonviral minicircle vector for deriving human iPS cells

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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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Figure 1: Generation of iPSCs with minicircle vector.
Figure 2: Pluripotency of mc-iPSCs.

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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

F.J., K.D.W., N.S., R.C.R., M.A.K., M.T.L. and J.C.W. conceived and designed the experiments. F.J., K.D.W., N.S., D.M.G., M.H., Z.Y.C., Z.L. and N.J.P. performed the experiments. F.J., K.D.W. and J.C.W. wrote the paper.

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

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–9 and Supplementary Tables 1–4 (PDF 615 kb)

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). (MOV 2062 kb)

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