Abstract
The production of lentiviral vectors (LVs) in human embryonic kidney 293 (HEK293) cells using serum-free medium in a suspension culture for the transduction of chimeric antigen receptor T-cells (CAR-T) can be achieved by different methods. This chapter describes LV production by transient transfection, induction of stable packaging cell lines, and induction of stable producer cell lines.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Naldini L (2015) Gene therapy returns to centre stage. Nature 526(7573):351–360
Escors D, Breckpot K (2010) Lentiviral vectors in gene therapy: their current status and future potential. Arch Immunol Ther Exp 58(2):107–119
Sharon D, Kamen A (2018) Advancements in the design and scalable production of viral gene transfer vectors. Biotechnol Bioeng 115(1):25–40
Naldini L, Trono D, Verma IM (2016) Lentiviral vectors, two decades later. Science 353(6304):1101–1102
Merten OW, Hebben M, Bovolenta C (2016) Production of lentiviral vectors. Mol Ther Methods Clin Dev 3:16017
Broussau S, Jabbour N, Lachapelle G et al (2008) Inducible packaging cells for large-scale production of lentiviral vectors in serum-free suspension culture. Mol Ther 16(3):500–507
Manceur AP, Kim H, Misic V et al (2017) Scalable lentiviral vector production using stable HEK293SF producer cell lines. Hum Gene Ther Methods 28(6):330–339
Ansorge S, Lanthier S, Transfiguracion J et al (2009) Development of a scalable process for high-yield lentiviral vector production by transient transfection of HEK293 suspension cultures. J Gene Med 11(10):868–876
Gelinas JF, Davies LA, Gill DR et al (2017) Assessment of selected media supplements to improve F/HN lentiviral vector production yields. Sci Rep 7(1):10198
Ansorge S, Henry O, Kamen A (2010) Recent progress in lentiviral vector mass production. Biochem Eng J 48(3):362–377
Logan AC, Nightingale SJ, Haas DL et al (2004) Factors influencing the titer and infectivity of lentiviral vectors. Hum Gene Ther 15(10):976–988
McCarron A, Donnelley M, McIntyre C et al (2016) Challenges of up-scaling lentivirus production and processing. J Biotechnol 240:23–30
Wang Y, Bergelson S, Feschenko M (2018) Determination of Lentiviral Infectious Titer by a Novel Droplet Digital PCR Method. Hum Gene Ther Methods 29(2):96–103
Tomás HA, Rodrigues AF, Carrondo MJT et al (2018) LentiPro26: novel stable cell lines for constitutive lentiviral vector production. Sci Rep 8(1):5271
Sanber KS, Knight SB, Stephen SL et al (2015) Construction of stable packaging cell lines for clinical lentiviral vector production. Sci Rep 5:9021
Acknowledgments
The authors would like to acknowledge Rénald Gilbert, Sven Ansorge, and their respective teams at NRC for providing HEK293 packaging and producer cell lines to support the research program with lentiviral vectors as well as funding from the Canada Research Chair (CRC-2403940) and Canadian Foundation for Innovation (CFI-32904).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Do Minh, A., Tran, M.Y., Kamen, A.A. (2020). Lentiviral Vector Production in Suspension Culture Using Serum-Free Medium for the Transduction of CAR-T Cells. In: Swiech, K., Malmegrim, K., Picanço-Castro, V. (eds) Chimeric Antigen Receptor T Cells. Methods in Molecular Biology, vol 2086. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0146-4_6
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0146-4_6
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0145-7
Online ISBN: 978-1-0716-0146-4
eBook Packages: Springer Protocols