Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences

By Junying Yu, Kejin Hu, Kimberley Smuga-Otto, Shulan Tian, Ron M. Stewart, I. Slukvin, James A. Thomson. Science, 2009

838 citations recorded by the source corpus.

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.

Subfields: Genetics, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/c343690f8a28815db1355a1db633a58cadff3ddd

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Genetics

In Molecular Biology

Citations in the corpus

Identification and classification of chromosomal aberrations in human induced pluripotent stem cells. (2010, methodology): “In the study of Yu et al. (2009), 16 HiPSC clones and subclones were determined to be normal by karyotype and a single small deletion in one subclone was identified by high-density CGH arrays (Yu et al., 2009).”
Highly efficient reprogramming to pluripotency and directed differentiation of human cells using synthetic modified mRNA (2010, methodology): “…plasmid, episomal, and adenovirus vectors (Chang et al., 2009; Kaji et al., 2009; Okita et al., 2008; Stadtfeld et al., 2008; Woltjen et al., 2009; Yu et al., 2009). iPSCs have also been derived with two DNA-free methods: serial protein transduction with recombinant proteins incorporating…”
Senescence impairs successful reprogramming to pluripotent stem cells. (2009, methodology): “Human integration-free iPS cells were later generated by adenoviral infection (Zhou and Freed, 2009), transfection with self-replicating episomal vectors (Yu et al., 2009) or with polycistronic minicircle vectors (Jia et al., 2010).”
Induced pluripotency: history, mechanisms, and applications. (2010, background): “…and Sendai virus (Fusaki et al. 2009), as well as with polycistronic minicircle vectors (Jia et al. 2010) and self-replicating selectable episomes (Yu et al. 2009), albeit the latter system required the simultaneous overexpression of additional factors, including another potent oncogene (Oct4,…”