Dynamics of magnetic lipoplexes studied by single particle tracking in living cells.
Magnetofection, gene delivery under the influence of a magnetic field, is a technique to increase transfection efficiency by enforcing gene vector contact with a target cell. Mechanisms of magnetic lipoplex internalization and intracellular details of magnetofection are still unknown. In this study, cellular dynamics of magnetic lipoplexes were examined in real time by means of highly sensitive dual-color fluorescence microscopy. Single particle tracking of magnetic lipoplexes provided trajectories representing the movement of the lipoplexes during internalization and subsequent intracellular processes. Magnetic lipoplexes show a three-phase behavior similar to polyplexes. During phase I lipoplexes are attached to the cell surface and show slow cooperative transport behavior. Phase II takes place inside the cell and was characterized by anomalous and confined diffusion. Phase III represented active transport along microtubules inside the cell. The majority of lipoplexes were internalized via endocytosis during phase I. On later time scales the formation of a perinuclear ring was observed. Persisting colocalization of fluid phase marker and lipoplexes after 24 h indicated slow endosomal release. In short, the internalization characteristics of magnetic lipoplexes are very similar to that of polyplexes. Furthermore our results suggest that the magnetic field induces an increased concentration of magnetic complexes on the cell surface resulting in higher transfection efficiency.
Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universität München, Butenandtstr. 5-13, D-81377 München, Germany.
A M
AM
Sauer
K G
KG
de Bruin
N
N
Ruthardt
O
O
Mykhaylyk
C
C
Plank
C
C
Bräuchle
D001692:Biological Transport; D045744:Cell Line, Tumor; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics*; D004337:Drug Carriers / Q000008:administration & dosage / Q000737:chemistry / Q000493:pharmacokinetics*; D004705:Endocytosis; D005290:Ferric Compounds / Q000737:chemistry; D005456:Fluorescent Dyes; D017930:Genes, Reporter; D006801:Humans; D008055:Lipids / Q000737:chemistry; D008156:Luciferases / Q000235:genetics; D008280:Magnetics; D008856:Microscopy, Fluorescence / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry; D014162:Transfection / Q000379:methods*
D004337:Drug Carriers; D005290:Ferric Compounds; D005456:Fluorescent Dyes; D008055:Lipids; C500466:metafectene; C000499:ferric oxide; D004247:DNA; D008156:Luciferases