Title of article :
In vitro labeling of neural stem cells with poly-L-Lysine coated super paramagnetic nanoparticles for green fluorescent protein transfection
Author/Authors :
Albukhaty, Salim tarbiat modares university - department of nanobiotechnology, تهران, ايران , Naderi-Manesh, Hossein tarbiat modares university - department of nanobiotechnology, تهران, ايران , Tiraihi, Taki tarbiat modares university - Medical Sciences Faculty - Department of Anatomical Sciences, تهران, ايران , Tiraihi, Taki khatam al-anbia hospital - shefa neurosciences research center, ايران
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 0.75μg/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 with the transfected cells showed green fluorescence. Conclusion: Transfection with SPION is an efficient, non-viral gene transfere method.
Keywords :
Nanoparticles , Neural stem cells (nsc) , Transfection
Journal title :
Iranian Biomedical Journal(IBJ)
Journal title :
Iranian Biomedical Journal(IBJ)