Poloxamer synperonic F108 improves cellular transduction with lentiviral vectors

J Gene Med. 2012 Aug;14(8):549-60. doi: 10.1002/jgm.2653.

Abstract

Background: Although lentiviral transduction methods are widely used, their broader application is dependent upon the optimization of lentiviral transduction efficiency for a broad range of cell types. In the present study, we focus on the evaluation of two chemical classes with respect to their ability to increase lentiviral transduction without cytotoxicity.

Methods: We compared the activity of adjuvants that are already used for lentivirus delivery with that of novel adjuvants selected on the basis of their chemical and physical characteristics.

Results: The novel poloxamer synperonic F108 demonstrated superior characteristics for enhancing lentiviral transduction over the best-in-class polybrene-assisted transduction. The results revealed that poloxamer synperonic F108 exhibited the dual benefits of low toxicity and a high efficiency of lentiviral gene delivery into a range of different primary cell cultures. In the presence of poloxamer synperonic F108, cells showed an increased propidium dye influx indicating a re-organization of membrane microstructures accompanying lentivirus uptake. The administration of a mixture of poloxamer synperonic F108 with polybrene further enhanced lentiviral transduction rates.

Conclusions: The results obtained in the present study indicate that a contribution to efficiency is made by each adjuvant, with polybrene acting as a charge protector and poloxamer synperonic F108 as a membrane modulator. Therefore, poloxamer synperonic F108, either alone or in combination, can lead to the optimization of large-scale lentiviral transduction approaches.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects
  • Cell Membrane / drug effects
  • Cell Proliferation / drug effects
  • Flow Cytometry
  • Genes, Reporter
  • Genetic Vectors
  • HEK293 Cells
  • Humans
  • Lentivirus / genetics*
  • Leukocytes, Mononuclear / metabolism
  • Leukocytes, Mononuclear / virology
  • Luciferases / biosynthesis
  • Luciferases / genetics
  • Permeability
  • Polyamines / pharmacology
  • Polyamines / toxicity
  • Polyelectrolytes
  • Polyethylenes / pharmacology*
  • Polyethylenes / toxicity
  • Polypropylenes / pharmacology*
  • Polypropylenes / toxicity
  • Surface-Active Agents / pharmacology*
  • Surface-Active Agents / toxicity
  • Transduction, Genetic / methods*

Substances

  • Polyamines
  • Polyelectrolytes
  • Polyethylenes
  • Polypropylenes
  • Surface-Active Agents
  • polycations
  • UCON 50-HB-5100
  • Luciferases