Abstract
More than 10 years ago, we developed an efficient protocol for serum-free retroviral transduction of human hematopoietic stem cells derived from mobilized peripheral blood. After upscaling of the methodology, serum-free retroviral gibbon-ape leukemia virus (GALV) pseudotype PG13/LN vector supernatant produced under strict good manufacturing practice (GMP) conditions was used in the first clinical gene-marking trial in Germany. In this study, we analyzed the titer and transduction efficiency of this serum-free clinical-grade retroviral supernatant 10 years after production to evaluate the long-term stability. Long-term storage and transport on dry ice resulted in modestly decreased titers and levels of transduction efficiency in CD34+ cells ranging from 38.4 to 49.1%. We conclude that the stability of retroviral vectors in serum-free medium allows extended storage and distribution of approved clinical-grade retroviral vector stocks to distant sites in multicenter clinical trials.
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Acknowledgements
We would like to thank Dr Roland Mertelsmann, University Hospital, Freiburg for supporting this study, and CellGenix GmbH, Freiburg for the long-term storage of the GMP-produced serum-free γ-retroviral supernatant. This work was supported by the Bundesministerium für Bildung und Forschung, Grant 01KV9907, by the Deutsche Forschungsgemeinschaft DFG, Grants KA 976/5-1, KA 976/5-2, Ka976/5-3 and Ka976/5-4, and by the Deutsche Krebshilfe Grants 10-1860-GI I and 107217.
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Herbst, F., Ball, C., Zavidij, O. et al. 10-Year stability of clinical-grade serum-free γ-retroviral vector-containing medium. Gene Ther 18, 210–212 (2011). https://doi.org/10.1038/gt.2010.126
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DOI: https://doi.org/10.1038/gt.2010.126