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Methods Mol Biol ; 480: 141-50, 2009.
Article in English | MEDLINE | ID: mdl-19085126

ABSTRACT

Transfer of exogenous material into the cytosol of cells is one of the main challenges in drug delivery. We present a novel physical approach for efficient incorporation of macromolecules into living cells, based on exposing them to a train of unipolar electric field pulses, possessing much lower amplitude than used for electroporation. The exposure of cells to a low electric field (LEF) alters the cell surface, leading to enhanced adsorption of macromolecules and their subsequent uptake by stimulated endocytosis. The macromolecules are initially encapsulated in membrane vesicles and then, at a later stage, are released into the cytosol and interact with intracellular targets. The uptake of fluorescently labeled macromolecules is monitored using confocal microscopy and flow cytometry. The biological activities of the incorporated macromolecules are determined by biochemical methods.


Subject(s)
Electroporation/methods , Endocytosis , Macromolecular Substances/administration & dosage , Animals , COS Cells , Chlorocebus aethiops , Flow Cytometry , Fluorescent Dyes , Microscopy, Confocal , Microscopy, Fluorescence , Molecular Probes
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