High-Titer Production of HIV-Based Lentiviral Vectors in Roller Bottles for Gene and Cell Therapy

  • Hazal Banu Olgun
  • Hale M. Tasyurek
  • Ahter Dilsad Sanlioglu
  • Salih SanliogluEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1879)


Lentiviral vectors are becoming preferred vectors of choice for clinical gene therapy trials due to their safety, efficacy, and the long-term gene expression they provide. Although the efficacy of lentiviral vectors is mainly predetermined by the therapeutic genes they carry, they must be produced at high titers to exert therapeutic benefit for in vivo applications. Thus, there is need for practical, robust, and scalable viral vector production methods applicable to any laboratory setting. Here, we describe a practical lentiviral production technique in roller bottles yielding high-titer third-generation lentiviral vectors useful for in vivo gene transfer applications. CaPO4-mediated transient transfection protocol involving the use of a transfer vector and three different packaging plasmids is employed to generate lentivectors in roller bottles. Following clearance of cellular debris via low-speed centrifugation and filtration, virus is concentrated by high-speed ultracentrifugation over sucrose cushion.


Gene therapy Lentivirus Roller bottles 



This study is supported by grants from Akdeniz University Scientific Research Administration Division (TYL-2015-1027) and the Scientific and Technological Research Council of Turkey (TUBITAK-112S114).


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Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  • Hazal Banu Olgun
    • 1
  • Hale M. Tasyurek
    • 1
  • Ahter Dilsad Sanlioglu
    • 1
  • Salih Sanlioglu
    • 1
    Email author
  1. 1.Human Gene and Cell Therapy Center of Akdeniz University HospitalsAntalyaTurkey

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