Cell internalization of magnetic nanoparticles using transfection agents.
Transfection agent (TFA)-induced magnetic cell labeling with Feridex IV is an attractive method of loading cells because it employs a pharmaceutical source of iron oxide. Although attractive, the method has two significant drawbacks. First, it requires mixing positively charged transfection agents and negatively charged magnetic nanoparticles, and the resulting loss of nanoparticle surface charge causes nanoparticle precipitation. Second, it can result in nanoparticle adsorption to the cell surface rather than internalization. Internalization of Feridex (and associated dextran) is important since dextran cell exterior can react with the antidextran antibodies, commonly present in human populations, and trigger an antibody-mediated cytotoxicity. Here we employed three assays for selecting Feridex/TFA mixtures to minimize nanoparticle precipitation and surface adsorption: (1) an assay for precipitation or stability (light scattering), (2) an assay for labeled cells (percentage of cells retained by a magnetic filter), and (3) an antidextran-based assay for nanoparticle internalization. Cells loaded with Feridex/protamine had internalized iron, whereas cells loaded with Feridex/Lipofectamine had surface-adsorbed iron. Optimal conditions for loading cells were 10 microg/Feridex and 3 microg/mL protamine sulfate. Conditions for loading cells with Feridex and a TFA need to be carefully selected to minimize nanoparticle precipitation and dextran adsorption to the cell surface.
Montet-Abou
Karin
K
Center for Molecular Imaging Research Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129, USA.
Montet
Xavier
X
Weissleder
Ralph
R
Josephson
Lee
L
D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D002477:Cells / Q000378:metabolism*; D003911:Dextrans; D052203:Ferrosoferric Oxide; D007501:Iron / Q000737:chemistry / Q000378:metabolism*; D058185:Magnetite Nanoparticles; D053768:Metal Nanoparticles; D051379:Mice; D010087:Oxides / Q000737:chemistry / Q000378:metabolism*; D010539:Permeability; D011107:Polylysine / Q000378:metabolism / Q000633:toxicity; D011479:Protamines / Q000378:metabolism*; D014162:Transfection
D003911:Dextrans; D058185:Magnetite Nanoparticles; D010087:Oxides; D011479:Protamines; D011107:Polylysine; D007501:Iron; C065507:ferumoxides; D052203:Ferrosoferric Oxide