Adenoviral Gene Delivery Can Reprogram Human Fibroblasts to Induced Pluripotent Stem Cells§

Authors

  • Wenbo Zhou,

    Corresponding author
    1. Departments of Medicine, Pharmacology, and Neurosurgery and the Neuroscience Program, University of Colorado Denver, School of Medicine, Aurora, Colorado, USA
    • Division of Clinical Pharmacology and Toxicology, Mailstop C-237, University of Colorado Denver, 12700 E. 19th Avenue, Aurora, Colorado 80045, USA
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    • Telephone: 303-724-6000; Fax: 303-724-6012

  • Curt R. Freed

    1. Departments of Medicine, Pharmacology, and Neurosurgery and the Neuroscience Program, University of Colorado Denver, School of Medicine, Aurora, Colorado, USA
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    • Telephone: 303-724-6000; Fax: 303-724-6012;


  • Author contributions: W.Z.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing; C.R.F.: conception and design, data analysis and interpretation, manuscript writing.

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

  • §

    First published online in STEM CELLS EXPRESS 2009.

Abstract

Mouse and human fibroblasts have been transformed into induced pluripotent stem (iPS) cells by retroviral transduction or plasmid transfection with four genes. Unfortunately, viral and plasmid DNA incorporation into chromosomes can lead to disruption of gene transcription and malignant transformation. Tumor formation has been found in offspring of mice generated from blastocysts made mosaic with iPS cells. To proceed with iPS cells for human therapy, reprogramming should be done with transient gene expression. Recently, adenoviral vectors have been used to produce mouse iPS cells without viral integration. Here, we report the successful creation of human iPS cells from embryonic fibroblasts using adenoviral vectors expressing c-Myc, Klf4, Oct4, and Sox2. After screening 12 colonies, three stable iPS cell lines were established. Each cell line showed human embryonic stem cell morphology and surface markers. Southern blots and polymerase chain reaction demonstrated that there was no viral DNA integration into iPS cells. Fingerprinting and karyotype analysis confirmed that these iPS cell lines are derived from the parent human fibroblasts. The three human iPS cell lines can differentiate to all three germ layers in vitro, including dopaminergic neurons. After s.c. injection into nonobese diabetic–severe combined immunodeficient mice, each human iPS line produced teratomas within 5 weeks postimplantation. We conclude that adenoviral vectors can reprogram human fibroblasts to pluripotent stem cells for use in individualized cell therapy without the risk for viral or oncogene incorporation. STEM CELLS 2009;27:2667–2674

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