Insights into the mechanism of magnetofection using PEI-based magnetofectins for gene transfer.
BACKGROUND
Gene delivery by the use of magnetic forces, so-called magnetofection, has been shown to enhance transfection efficiency of viral and non-viral systems up to several-hundred-fold. For this purpose gene carriers, such as polyethylenimine (PEI), are associated with superparamagnetic nanoparticles and complexed with plasmid DNA. Gene delivery is targeted by the application of a magnetic field.


METHODS
To investigate the underlying mechanism, we studied the impact of the applied magnetic field on the transfection process of PEI-coated superparamagnetic iron oxide gene vectors (magnetofectins) using various cell lines. In particular, we addressed the question whether accelerated sedimentation of magnetofectins is the driving force or if the magnetic field itself directly influences the endocytic processing of the magnetofectins. The cellular uptake mechanism of magnetofectins was studied by electron microscopy and transfection experiments in the presence of various inhibitors that operate at different steps of endocytosis.


RESULTS
In this study we could show that cellular uptake of magnetofectins proceeds obviously by endocytosis. Cellular uptake of magnetofectins behaves almost analogously as compared with PEI polyplexes. Besides unspecific endocytosis, apparently clathrin-dependent as well as caveolae-mediated endocytic uptake is involved.


CONCLUSIONS
The magnetic field itself does not alter the uptake mechanism of magnetofectins. Obviously, the magnetic forces lead to an accelerated sedimentation of magnetofectins on the cell surface and do not directly affect the endocytic uptake mechanism. So further improvement of magnetic field application could lead to efficient targeting of gene expression into the desired organ and tissue in vivo.
Department of Pediatrics, Ludwig-Maximilians-University, 80337 Munich, Germany.
Stephanie
S
Huth
James
J
Lausier
Soeren W
SW
Gersting
Carsten
C
Rudolph
Christian
C
Plank
Ulrich
U
Welsch
Joseph
J
Rosenecker
D000968:Antimycin A / Q000494:pharmacology; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D004705:Endocytosis; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008280:Magnetics; D008854:Microscopy, Electron; D010957:Plasmids; D011094:Polyethyleneimine / Q000737:chemistry*; D014162:Transfection / Q000379:methods*
D000968:Antimycin A; D011094:Polyethyleneimine; D008156:Luciferases