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The FBMD-1 stroma cell line secretes a unique moiety which can increase retroviral transduction of lineage-committed and primitive human peripheral blood progenitor cells

Cancer Gene Therapy volume 8, pages 440–449 (2001)Cite this article

Abstract

Peripheral blood progenitor cells are a prime target for gene therapy approaches. As recent data point to the relevance of soluble stroma factors for the efficient transduction of progenitor cells, we tested the stroma-conditioned medium (SCM) of the two cell lines FBMD-1 and L88/5 as well as desulfated and O-sulfated heparin (HS dS and HS OS) for their effect on transduction of peripheral blood progenitor cells. We transduced CD34 + cells of nine tumor patients with the retroviral SF-MDR vector containing the human multidrug resistance 1 ( MDR 1) gene under serum-free conditions on the fibronectin fragment CH-296 with or without SCM. Provirus-specific polymerase chain reaction showed a median 1.6-fold higher integration rate of the transgene into committed progenitor cells for the group with added FBMD-1 SCM ( P =.008). This was maintained after 2 ( P =.02) and, as a trend, after 5 weeks of stroma-dependent long-term culture. We found a median 1.5-fold increase in rhodamine-123 (Rh-123) exclusion in myeloid lineage-committed progeny cells following transduction in the presence of FBMD-1 SCM ( P =.0004). After 2 or 5 weeks of long-term culture, a significantly higher proportion of Rh-123 dull cells could still be detected in the FBMD-1 SCM transduction group ( P =.003 and P =.04, respectively). L88/5 SCM or HS OS or HS dS was not effective as supplement for improving gene transfer. The FBMD-1 stroma cell line appears to secrete a unique moiety, which can increase retroviral transduction of lineage-committed and primitive progenitor cells. The FBMD-1 stroma activity is not attributable to heparan sulfate. Cancer Gene Therapy (2001) 8, 440–449

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Acknowledgements

The authors thank Sigrid Heil, Hans Jürgen Engel, and Bernhard Berkus for their expert technical help. We also thank the Stem Cell Transplantation team of the Department of Internal Medicine V, University of Heidelberg, for their help with collecting the cell samples of the patients. The FBMD-1 cell line was kindly provided by R. Ploemacher, Institute of Hematology, Erasmus University, Rotterdam, the Netherlands. The SF-MDR vector was kindly made available by C. Baum, Heinrich-Pette-Institut, Hamburg, Germany. Flt-3 ligand was a kind gift of D. Williams, PhD, Immunex, Seattle, WA. The L88/5 cell line was kindly provided by K. Thalmeier, GSF-Forschungszentrum für Umwelt und Gesundheit, Munich, Germany. A. Benner of the Central Unit Biostatistics at the German Cancer Research Center gave invaluable help on the statistical calculations. We thank K. Wermann for helpful comments on the manuscript. This work was supported, in part, by Grant 10-1294-Ze2 of the “Deutsche Krebshilfe/Dr. Mildred Scheel Stiftung.”

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  1. German Cancer Research Center, Research Program Diagnostics and Experimental Therapy, Heidelberg, D0200, Germany

    Eike C Buss, Bernd Schiedlmeier & W Jens Zeller

  2. Department of Internal Medicine V, University of Heidelberg, Heidelberg, Germany

    Anthony D Ho & Stefan Fruehauf

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  1. Eike C Buss
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Correspondence to Stefan Fruehauf.

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Buss, E., Schiedlmeier, B., Ho, A. et al. The FBMD-1 stroma cell line secretes a unique moiety which can increase retroviral transduction of lineage-committed and primitive human peripheral blood progenitor cells. Cancer Gene Ther 8, 440–449 (2001). https://doi.org/10.1038/sj.cgt.7700324

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