The transfection of multipotent neural precursor/stem cell transplant populations with magnetic nanoparticles

Part I: Minicircle vector technology limits DNA size restrictions on ex vivo gene delivery using nanoparticle vectors: Overcoming a translational barrier in neural stem cell therapy

“…We recently reported technically simple, rapid and safe magnetofection protocols for genetic modification of major neural transplant types including NSCs, with oscillating magnetic fields always outperforming static fields [21][22][23][24][25]. MNPs have emerged strongly as a class of advanced functional materials for neuro-regeneration, serving as 'multifunctional tools' for cell therapy given additional applications for non-invasive cell imaging and magnetic cell targeting, so it is clear that this non-viral gene transfer method offers significant benefits for clinical translation [26].…”

“…Double transduction/transfection is routinely used for many viral as well as non-viral genetic modification approaches such as lipid-mediated gene delivery and electroporation in order to enhance transfection efficiency and to prolong gene expression [37][38][39]. We previously showed that multifection of NSCs (cultured as 3-D neurospheres) resulted in higher transfection levels (~27%) compared to single transfection (~13%) with no effect on cell physiology and health [22]. NSC monolayers multifected here with mC-GFP showed significantly higher transfection (see Fig.…”

“…Notably, a sustained yet tapered profile of therapeutic biomolecule expression may be beneficial as it mirrors the temporal pattern of molecular expression profiles that are associated with regenerative processes in pathology sites [22]. Indeed, it has been suggested that…”

Part II: Functional delivery of a neurotherapeutic gene to neural stem cells using minicircle DNA and nanoparticles: Translational advantages for regenerative neurology

“…4 with oscillating magnetic fields can offer a safe solution for gene delivery in neural cells [15][16][17][18][19][20][21][22][23] but a major challenge encountered has been reduced transfection associated with increased size of therapeutic plasmids [23][24][25][26][27] (discussed in detail in Part I).…”

Multimodal Magnetic Core–Shell Nanoparticles for Effective Stem‐Cell Differentiation and Imaging

“…To see whether we could increase transfection efficiency further, we carried out repeated transfections of the same cell culture (a technique known as multifection [18] ) using our MCNP/siGFP constructs and compared the gene-silencing efficiency achieved with multifection to that of a single transfection. We found that we were able to further improve the gene silencing efficiency from 55% (single transfection of MCNP/siGFP) to 65% (multifection of MCNP/siGFP) (See supporting information, Figure S10a).…”