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.
Gene Expression Omnibus
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.).
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).