The labeling of cationic iron oxide nanoparticle-resistant hepatocellular carcinoma cells using targeted magnetoliposomes.
The in vitro labeling of cultured cells with nanomaterials is a frequent practice but the efficiency, specificity and cytotoxicity of labeling specific cell types using targeted nanoparticles has only rarely been investigated. In the present work, functionalized anionic lipid-coated iron oxide cores (magnetoliposomes (MLs)) bearing galactose moieties were used for the specific labeling of asialoglycoprotein receptor 1 (ASGPR-1)-expressing HepG2 cells. The optimal number of galactose moieties per particle (± 26) was determined and uptake efficiency was compared with galactose-lacking anionic and cationic MLs. Using a blocking assay with free galactose, electron microscopy and co-cultures of HepG2 and non-ASGPR-1 expressing C17.2 cells, the specificity of the particles for the ASGPR-1 receptor was demonstrated. The intracellular localization of the galactose-bearing MLs was further verified by confocal microscopy. The non-toxic ML concentration was determined to be 400 μg Fe/ml. Finally, the use of these MLs for visualization of labelled cells by magnetic resonance imaging (MRI) was demonstrated. The data show a high uptake and specificity of the galactose-bearing MLs, whereas the cationic MLs remain primarily surface-associated. Thus, targeted MLs offer a successful alternative for cell labeling when cationic particles fail to be efficiently internalized.
Lab of BioNanoColloids, Interdisciplinary Research Centre, Katholieke Universiteit Leuven - Campus Kortrijk, B-8500 Kortrijk, Belgium.
Stefaan J H
SJ
Soenen
Alain R
AR
Brisson
Eveline
E
Jonckheere
Nele
N
Nuytten
Sisareuth
S
Tan
Uwe
U
Himmelreich
Marcel
M
De Cuyper
D000818:Animals; D002118:Calcium / Q000378:metabolism; D006528:Carcinoma, Hepatocellular; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D005290:Ferric Compounds / Q000737:chemistry* / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D008081:Liposomes / Q000737:chemistry*; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*; D014168:Transferrin / Q000378:metabolism
D005290:Ferric Compounds; D008081:Liposomes; D017382:Reactive Oxygen Species; D014168:Transferrin; C000499:ferric oxide; D002118:Calcium