• Influence of Amplitude of Oscillating Magnetic Fields on Magnetic Nanoparticle-Mediated Gene Transfer to Astrocytes

      Tickle, Jacqueline A.; Jenkins, Stuart I.; Pickard, Mark R.; Chari, Divya M.; Keele University, United Kingdom (World Scientific, 2014-08-07)
      Functionalized magnetic nanoparticles (MNPs) are emerging as a major nanoplatform for regenerative neurology, particularly as transfection agents for gene delivery. Magnetic assistive technology, particularly the recent innovation of applied oscillating magnetic fields, can significantly enhance MNP-mediated gene transfer to neural cells. While transfection efficiency varies with oscillation frequency in various neural cell types, the influence of oscillation amplitude has not yet been investigated. We have addressed this issue using cortical astrocytes that were transfected using MNPs functionalized with plasmid encoding a reporter protein. Cells were exposed to a range of oscillation amplitudes (100–1000 μm), using a fixed oscillation frequency of 1 Hz. No significant differences were found in the proportions of transfected cells at the amplitudes tested, but GFP-related optical density measurements (indicative of reporter protein expression) were significantly enhanced at 200 μm. Safety data show no amplitude-dependent toxicity. Our data suggest that the amplitude of oscillating magnetic fields influences MNP-mediated transfection, and a tailored combination of amplitude and frequency may further enhance transgene expression. Systematic testing of these parameters in different neural subtypes will enable the development of a database of neuro-magnetofection protocols — an area of nanotechnology research where little information currently exists.
    • Using magnetic nanoparticles for gene transfer to neural stem cells: stem cell propagation method influences outcomes

      Pickard, Mark R.; Adams, Christopher F.; Barraud, Perrine; Chari, Divya M.; Keele University, United Kingdom; University of Cambridge, United Kingdom (Multidisciplinary Digital Publishing Institute, 2015-04-24)
      Genetically engineered neural stem cell (NSC) transplants offer a key strategy to augment neural repair by releasing therapeutic biomolecules into injury sites. Genetic modification of NSCs is heavily reliant on viral vectors but cytotoxic effects have prompted development of non-viral alternatives, such as magnetic nanoparticle (MNPs). NSCs are propagated in laboratories as either 3-D suspension "neurospheres" or 2-D adherent "monolayers". MNPs deployed with oscillating magnetic fields ("magnetofection technology") mediate effective gene transfer to neurospheres but the efficacy of this approach for monolayers is unknown. It is important to address this issue as oscillating magnetic fields dramatically enhance MNP-based transfection in transplant cells (e.g., astrocytes and oligodendrocyte precursors) propagated as monolayers. We report for the first time that oscillating magnetic fields enhanced MNP-based transfection with reporter and functional (basic fibroblast growth factor; FGF2) genes in monolayer cultures yielding high transfection versus neurospheres. Transfected NSCs showed high viability and could re-form neurospheres, which is important as neurospheres yield higher post-transplantation viability versus monolayer cells. Our results demonstrate that the combination of oscillating magnetic fields and a monolayer format yields the highest efficacy for MNP-mediated gene transfer to NSCs, offering a viable non-viral alternative for genetic modification of this important neural cell transplant population.