Induced Pluripotent Stem Cell Generation Using a Single Lentiviral Stem Cell Cassette §

Authors

  • Cesar A. Sommer,

    1. Section of Gastroenterology, Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, USA
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  • Matthias Stadtfeld,

    1. Massachusetts General Hospital Cancer Center and Center for Regenerative Medicine, Boston, Massachusetts, USA
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  • George J. Murphy,

    1. Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA
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  • Konrad Hochedlinger,

    1. Massachusetts General Hospital Cancer Center and Center for Regenerative Medicine, Boston, Massachusetts, USA
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  • Darrell N. Kotton,

    1. Boston University Pulmonary Center, Boston University School of Medicine, Boston, Massachusetts, USA
    2. Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, USA
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  • Gustavo Mostoslavsky

    Corresponding author
    1. Section of Gastroenterology, Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, USA
    • Section of Gastroenterology, Department of Medicine, Boston University School of Medicine, 650 Albany Street X-513, Boston, Massachusetts 02118, USA

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    • Telephone: 617-638-6532; Fax: 617-638-7785


  • First published online in STEM CELLS Express December 18, 2008.

  • Disclosure of potential conflicts of interest is found at the end of this article.

  • §

    Author contributions: C.A.S.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing; M.S. and G.J.M.: collection and assembly of data; K.H.: conception and design, data analysis and interpretation; D.N.K.: conception and design, data analysis and interpretation, manuscript writing; G.M.: conception and design, data analysis and interpretation, manuscript writing, final approval of manuscript. M.S. and G.J.M. contributed equally to this work.

Abstract

Induced pluripotent stem (iPS) cells can be generated using retroviral vectors expressing Oct4, Klf4, Sox2, and cMyc. Most prior studies have required multiple retroviral vectors for reprogramming, resulting in high numbers of genomic integrations in iPS cells and limiting their use for therapeutic applications. Here we describe the use of a single lentiviral vector expressing a “stem cell cassette” composed of the four transcription factors and a combination of 2A peptide and internal ribosome entry site technology, generating iPS cells from postnatal fibroblasts. iPS cells generated in this manner display embryonic stem cell-like morphology, express stem cell markers, and exhibit in vivo pluripotency, as evidenced by their ability to differentiate in teratoma assays and their robust contribution to mouse chimeras. Combining all factors into a single transcript achieves the most efficient reprogramming system to date and allows derivation of iPS cells with a single viral integration. The use of a single lentiviral vector for reprogramming represents a powerful laboratory tool and a significant step toward the application of iPS technology for clinical purposes. STEM CELLS 2009;27:543–549

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