Fibroblast-Derived Induced Pluripotent Stem Cells Show No Common Retroviral Vector Insertions (original) (raw)

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Differentiation and Cancer, Center for Genomic Regulation and Pompeu Fabra University

, Barcelona,

Spain

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Cancer Center and Center for Regenerative Medicine, Massachusetts General Hospital, Harvard Stem Cell Institute

, Boston, Massachusetts,

USA

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Differentiation and Cancer, Center for Genomic Regulation and Pompeu Fabra University

, Barcelona,

Spain

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,

Cancer Center and Center for Regenerative Medicine, Massachusetts General Hospital, Harvard Stem Cell Institute

, Boston, Massachusetts,

USA

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,

Differentiation and Cancer, Center for Genomic Regulation and Pompeu Fabra University

, Barcelona,

Spain

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Bioinformatics Program, Center for Genomic Regulation and Pompeu Fabra University

, Barcelona,

Spain

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Bioinformatics Program, Center for Genomic Regulation and Pompeu Fabra University

, Barcelona,

Spain

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Cancer Center and Center for Regenerative Medicine, Massachusetts General Hospital, Harvard Stem Cell Institute

, Boston, Massachusetts,

USA

Correspondence: Konrad Hochedlinger, Ph.D., Center for Regenerative Medicine and Cancer Center, Massachusetts General Hospital, 185 Cambridge Street, CPZN 4242, Boston, Massachusetts 02114, USA. Telephone: 617-643-2075; Fax: 617-643-3170; e-mail: khochedlinger@helix.mgh.harvard.edu

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Differentiation and Cancer, Center for Genomic Regulation and Pompeu Fabra University

, Barcelona,

Spain

Institució Catalana de Recerca i Estudis Avançats

Correspondence: Thomas Graf, Ph.D., Centre for Genomic Regulation (CRG), C/Dr. Aiguader, 88, 08003 Barcelona, Spain. Telephone: 34-93-316-01-27; Fax: 34-93-316-00-99; e-mail: thomas.graf@crg.es

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Accepted:

04 October 2008

Published:

02 February 2009

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Florencio Varas, Matthias Stadtfeld, Luisa de Andres-Aguayo, Nimet Maherali, Alessandro di Tullio, Lorena Pantano, Cedric Notredame, Konrad Hochedlinger, Thomas Graf, Fibroblast-Derived Induced Pluripotent Stem Cells Show No Common Retroviral Vector Insertions, Stem Cells, Volume 27, Issue 2, February 2009, Pages 300–306, https://doi.org/10.1634/stemcells.2008-0696
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Abstract

Several laboratories have reported the reprogramming of mouse and human fibroblasts into pluripotent cells, using retroviruses carrying the Oct4, Sox2, Klf4, and c-Myc transcription factor genes. In these experiments the frequency of reprogramming was lower than 0.1% of the infected cells, raising the possibility that additional events are required to induce reprogramming, such as activation of genes triggered by retroviral insertions. We have therefore determined by ligation-mediated polymerase chain reaction (LM-PCR) the retroviral insertion sites in six induced pluripotent stem (iPS) cell clones derived from mouse fibroblasts. Seventy-nine insertion sites were assigned to a single mouse genome location. Thirty-five of these mapped to gene transcription units, whereas 29 insertions landed within 10 kilobases of transcription start sites. No common insertion site was detected among the iPS clones studied. Moreover, bioinformatics analyses revealed no enrichment of a specific gene function, network, or pathway among genes targeted by retroviral insertions. We conclude that Oct4, Sox2, Klf4, and c-Myc are sufficient to promote fibroblast-to-iPS cell reprogramming and propose that the observed low reprogramming frequencies may have alternative explanations.

Copyright © 2009 AlphaMed Press

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