In vitro labeling of neural stem cells with poly-L-lysine coated super paramagnetic nanoparticles for green fluorescent protein transfection

Iran Biomed J. 2013 Apr;17(2):71-6. doi: 10.6091/ibj.1114.2013.

Abstract

Background: The magnetic nanoparticle-based transfection method is a relatively new technique for delivery of functional genes to target tissues. We aimed to evaluate the transfection efficiency of rat neural stem cell (NSC) using poly-L-lysine hydrobromide (PLL)-coated super paramagnetic iron oxide nanoparticles (SPION).

Methods: The SPION was prepared and coated with PLL as transfection agent and the transfection efficiency was evaluated in rat NSC using enhanced green fluorescent protein-N1 plasmid containing GFP as a reporter gene. NSC was incubated for 24 h in cell culture media containing 25 µg/ml SPION and in different concentrations of PLL (0.25, 0.50, 0.75, 1 and 2 µg/ml). Cell viability was determined before and after transfection for every concentration using Trypan blue assay. Characterization of prepared uncoated (SPION) and coated (SPION-PLL) complexes were evaluated by a transmission electron microscope and the zeta potential.

Results: PLL at 0.75 μg/ml showed optimal results with 25 μg/ml SPION concentration compared with other PLL concentrations (0.25, 0.50, 1 and 2 μg/ml). The 18% efficiency of the transfected cells showed green fluorescence.

Conclusion: Transfection with SPION is an efficient, non-viral gene transfere method.

Keywords: Neural Stem cells; Nanoparticles; Transfection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Shape / drug effects
  • Cell Survival / drug effects
  • Dextrans
  • Green Fluorescent Proteins / metabolism*
  • Magnetite Nanoparticles
  • Nanoparticles / chemistry*
  • Nanoparticles / ultrastructure
  • Neural Stem Cells / cytology
  • Neural Stem Cells / drug effects
  • Neural Stem Cells / metabolism*
  • Polylysine / pharmacology*
  • Rats
  • Rats, Sprague-Dawley
  • Staining and Labeling*
  • Transfection*

Substances

  • Dextrans
  • Magnetite Nanoparticles
  • Green Fluorescent Proteins
  • Polylysine
  • ferumoxides