Open Access

Optimization of lentiviral vector production using polyethylenimine‑mediated transfection

  • Authors:
    • Yong Tang
    • Kenneth Garson
    • Li Li
    • Barbara C. Vanderhyden
  • View Affiliations

  • Published online on: November 7, 2014     https://doi.org/10.3892/ol.2014.2684
  • Pages: 55-62
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Abstract

The aim of the present study was to optimize the polyethylenimine (PEI)‑mediated transfection method in order to simplify the efficient production of lentiviral vectors (LvVs), and to compare the CaPO4‑ and PEI‑mediated transfection methods for producing LvVs. Different titration methods of LvV stocks, as well as different culture media, culture durations, cell densities and DNA quantities were compared to obtain an optimized procedure for the production of LvVs. Optimization of the production method for LvVs was achieved using PEI‑mediated transient transfections. Serum‑free Opti‑MEM® was used to directly produce LvVs that could be harvested 48 h after transfection. Furthermore, a cell density of 15x106 cells/10‑cm plate and a DNA concentration of 1X were selected for the optimum production of LvVs. The optimized LvV titration method was simple and direct; it involved LvVs carrying fluorescent reporters, which proved to be faster than the standard methods but equally as sensitive. In conclusion, a scalable process for production of LvVs by PEI‑mediated transfection was established and optimized. The optimized PEI‑mediated transfection method was easy to use, as well as providing greater reliability with a higher degree of reproducibility and consistency. Despite using less DNA, the PEI‑mediated transfection method resulted in viral titers that were the same as those achieved using the CaPO4‑mediated method.
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January-2015
Volume 9 Issue 1

Print ISSN: 1792-1074
Online ISSN:1792-1082

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Spandidos Publications style
Tang Y, Garson K, Li L and Vanderhyden BC: Optimization of lentiviral vector production using polyethylenimine‑mediated transfection. Oncol Lett 9: 55-62, 2015.
APA
Tang, Y., Garson, K., Li, L., & Vanderhyden, B.C. (2015). Optimization of lentiviral vector production using polyethylenimine‑mediated transfection. Oncology Letters, 9, 55-62. https://doi.org/10.3892/ol.2014.2684
MLA
Tang, Y., Garson, K., Li, L., Vanderhyden, B. C."Optimization of lentiviral vector production using polyethylenimine‑mediated transfection". Oncology Letters 9.1 (2015): 55-62.
Chicago
Tang, Y., Garson, K., Li, L., Vanderhyden, B. C."Optimization of lentiviral vector production using polyethylenimine‑mediated transfection". Oncology Letters 9, no. 1 (2015): 55-62. https://doi.org/10.3892/ol.2014.2684