10-year stability of clinical-grade serum-free γ-retroviral vector-containing medium.
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.
Department of Translational Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center, Im Neuenheimer Feld 350, Heidelberg, Germany.
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Herbst
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Ball
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Zavidij
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Fessler
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Schmidt
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Veelken
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von Kalle
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Glimm
D016895:Culture Media, Serum-Free; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells; D017989:Leukemia Virus, Gibbon Ape / Q000235:genetics*; D011309:Preservation, Biological; D013997:Time Factors; D014161:Transduction, Genetic
D016895:Culture Media, Serum-Free