Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences

Designer Stem Cells Despite their promise for use as disease models and in regenerative medicine, the generation of human-induced pluripotent stem (iPS) cells has been hindered by the integration of vector and transgenes in the host cell genome. Recent studies using the Cre/LoxP recombination strategy and the piggyBac transposon approach have approached this objective. However, Yu et al. (p. 797, published online 26 March) now show the derivation of human iPS cells from postnatal foreskin fibroblasts using the nonintegrating oriP/EBNA1-based episomal vectors. The resultant iPS cells show characteristics of human embryonic stem cells and are free of vector and transgenes. Human induced pluripotent stem cells can be generated without integration of exogenous DNA into their genomes. Reprogramming differentiated human cells to induced pluripotent stem (iPS) cells has applications in basic biology, drug development, and transplantation. Human iPS cell derivation previously required vectors that integrate into the genome, which can create mutations and limit the utility of the cells in both research and clinical applications. We describe the derivation of human iPS cells with the use of nonintegrating episomal vectors. After removal of the episome, iPS cells completely free of vector and transgene sequences are derived that are similar to human embryonic stem (ES) cells in proliferative and developmental potential. These results demonstrate that reprogramming human somatic cells does not require genomic integration or the continued presence of exogenous reprogramming factors and removes one obstacle to the clinical application of human iPS cells.

[1]  Wei Wang,et al.  piggyBac transposition reprograms fibroblasts to induced pluripotent stem cells , 2009, Nature.

[2]  Rudolf Jaenisch,et al.  Parkinson's Disease Patient-Derived Induced Pluripotent Stem Cells Free of Viral Reprogramming Factors , 2009, Cell.

[3]  C. Lorson,et al.  Induced pluripotent stem cells from a spinal muscular atrophy patient , 2009, Nature.

[4]  Shinya Yamanaka,et al.  Generation of Mouse Induced Pluripotent Stem Cells Without Viral Vectors , 2008, Science.

[5]  J. Utikal,et al.  Induced Pluripotent Stem Cells Generated Without Viral Integration , 2008, Science.

[6]  Sheng Ding,et al.  Induction of pluripotent stem cells from mouse embryonic fibroblasts by Oct4 and Klf4 with small-molecule compounds. , 2008, Cell stem cell.

[7]  A. Consiglio,et al.  Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes , 2008, Nature Biotechnology.

[8]  Wenjun Guo,et al.  Induction of pluripotent stem cells by defined factors is greatly improved by small-molecule compounds , 2008, Nature Biotechnology.

[9]  Jiuhong Kang,et al.  Enhanced efficiency of generating induced pluripotent stem (iPS) cells from human somatic cells by a combination of six transcription factors , 2008, Cell Research.

[10]  Shulan Tian,et al.  Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells , 2007, Science.

[11]  T. Ichisaka,et al.  Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors , 2007, Cell.

[12]  A. Nanbo,et al.  The coupling of synthesis and partitioning of EBV's plasmid replicon is revealed in live cells , 2007, The EMBO journal.

[13]  T. Ichisaka,et al.  Generation of germline-competent induced pluripotent stem cells , 2007, Nature.

[14]  S. Yamanaka,et al.  Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors , 2006, Cell.

[15]  刘金明,et al.  IL-13受体α2降低血吸虫病肉芽肿的炎症反应并延长宿主存活时间[英]/Mentink-Kane MM,Cheever AW,Thompson RW,et al//Proc Natl Acad Sci U S A , 2005 .

[16]  B. Sugden,et al.  Establishment of an oriP Replicon Is Dependent upon an Infrequent, Epigenetic Event , 2001, Molecular and Cellular Biology.

[17]  Robert A. Weinberg,et al.  Creation of human tumour cells with defined genetic elements , 1999, Nature.

[18]  J. Thomson,et al.  Embryonic stem cell lines derived from human blastocysts. , 1998, Science.

[19]  Gerard I. Evan,et al.  Induction of apoptosis in fibroblasts by c-myc protein , 1992, Cell.

[20]  J. Yates,et al.  Epstein-Barr virus-derived plasmids replicate only once per cell cycle and are not amplified after entry into cells , 1991, Journal of virology.

[21]  D. Reisman,et al.  A cis-acting element from the Epstein-Barr viral genome that permits stable replication of recombinant plasmids in latently infected cells. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[22]  L. Zon,et al.  Cell stem cell. , 2007, Cell stem cell.

[23]  J. Yates,et al.  Stable replication of plasmids derived from Epstein–Barr virus in various mammalian cells , 1985, Nature.