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Improved lentiviral transduction of human mesenchymal stem cells for therapeutic intervention in pancreatic cancer

Cancer Gene Therapy volume 15, pages 231–240 (2008)Cite this article

Abstract

Genetic modification of human bone marrow mesenchymal stem cells (MSC) is highly valuable for their exploitation in basic science and therapeutic applications, for example in cancer. We present here a new, fast and easy-to-use method to enrich a functional population of lentiviral (LV)-transduced MSC expressing enhanced green fluorescent protein (eGFP). We replaced the eGFP gene by a fusion gene of puromycin acetyltransferase and eGFP. Upon LV gene transfer and puromycin selection, we quickly obtained a pure transduced MSC population, in which growth, differentiation capacity and migration preferences were not compromised. Furthermore, we are the first to report the migration velocity of MSC among which 30% were moving and velocity of about 15 μm h−1 was not altered by LV transduction. Manipulated MSC underwent senescence one passage earlier than non-transduced cells, suggesting the use for therapeutic intervention in early passage numbers. Upon tail vein application in nude mice, the majority of LV-transduced MSC could be detected in human orthotopic pancreatic tumor xenografts and to a minor extent in mouse liver, kidney and lung. Together, LV transduction of genes to MSC followed by puromycin selection is a powerful tool for basic research and improves the therapeutic prospects of MSC as vehicles in gene therapy.

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Acknowledgements

We thank Dr van Parijs for providing the lentiviral vector pLL3.7 and K Hexel for help in FACS sorting. This study was supported by grants from the Tumorzentrum Heidelberg/Mannheim, Deutsche Krebshilfe and BMBF.

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Author notes

  1. G Kallifatidis, B M Beckermann, A Groth and M Schubert: These four authors contributed equally to this work.

  2. P Büchler and I Herr: These two authors contributed equally to this work.

Authors and Affiliations

  1. Molecular OncoSurgery, University of Heidelberg and German Cancer Research Center, Heidelberg, Germany

    G Kallifatidis, B M Beckermann, A Groth, A Apel, T Wenger, J Mattern & I Herr

  2. Department of General Surgery, University of Heidelberg, Heidelberg, Germany

    G Kallifatidis, B M Beckermann, A Groth, A Apel, A Khamidjanov, E Ryschich, M W Büchler, H Friess, P Büchler & I Herr

  3. Department of Medicine V, University of Heidelberg, Heidelberg, Germany

    M Schubert, W Wagner, A Diehlmann, R Saffrich, U Krause, V Eckstein & A D Ho

  4. Centre d'Immunologie de Marseille-Luminy, Marseille, France

    T Wenger

  5. Department of Physiology and Pathophysiology, University of Heidelberg, Heidelberg, Germany

    W Wagner

  6. Department of Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany

    M Chai & G Schütz

  7. Department of Experimental Surgery, University of Heidelberg, Heidelberg, Germany

    M M Gebhard, H Friess & P Büchler

  8. Department of Surgery, Klinik rechts der Isar, Technical University of Munich, Munich, Germany

    M M Gebhard, H Friess & P Büchler

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  1. G Kallifatidis
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Correspondence to I Herr.

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Supplementary Information accompanies the paper on Cancer Gene Therapy website (http://www.nature.com/cgt)

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Kallifatidis, G., Beckermann, B., Groth, A. et al. Improved lentiviral transduction of human mesenchymal stem cells for therapeutic intervention in pancreatic cancer. Cancer Gene Ther 15, 231–240 (2008). https://doi.org/10.1038/sj.cgt.7701097

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