Co-localization of fluorescent labeled lipid nanoparticles with specifically tagged subcellular compartments by single particle tracking at low nanoparticle to cell ratios

Autor: Francis C. Szoka, Matthew Tiffany
Rok vydání: 2016
Předmět:
0301 basic medicine
Endosome
media_common.quotation_subject
Pharmaceutical Science
02 engineering and technology
Polyethylene glycol
Endocytosis
Polyethylene Glycols
Fatty Acids
Monounsaturated
Propanolamines
HeLa
03 medical and health sciences
chemistry.chemical_compound
Drug Delivery Systems
Lysosome
medicine
Humans
RNA
Small Interfering
Luciferases
Internalization
Triglycerides
Fluorescent Dyes
media_common
biology
Chemistry
Phosphatidylethanolamines
technology
industry
and agriculture
Biological Transport
Lipid metabolism
Lipid Metabolism
021001 nanoscience & nanotechnology
biology.organism_classification
Lipids
In vitro
Quaternary Ammonium Compounds
Cholesterol
030104 developmental biology
medicine.anatomical_structure
Biochemistry
Biophysics
Nanoparticles
lipids (amino acids
peptides
and proteins)
0210 nano-technology
HeLa Cells
Zdroj: Journal of Drug Targeting. 24:857-864
ISSN: 1029-2330
1061-186X
DOI: 10.1080/1061186x.2016.1233976
Popis: We utilized quantitative high-resolution single particle tracking to study the internalization and endosomal sorting of lipid nanoparticles (LNPs) by HeLa cells in vitro to gain a better understanding of how cells process LNPs that are used for siRNA delivery. We compared the trafficking of three formulations that have been demonstrated to deliver siRNA into cells. They were composed of either a tritratable anionic lipid, formulation of cholesterol hemisuccinate (CHEMS), or a titratatable cationic lipid formulation of 1,2-dilinoleyloxy-3-dimethylaminopropane (DLinDMA) or a non-titratable cationic formulation lipid formulation of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP). They also contained either a substantial percentage of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) or cholesterol and 5 mole percent 1,2-dimyristoyl-sn-glycerol-[methoxy(polyethylene glycol)-2000 (PEG-DMG). We optically measured the endosomal pH experienced by individual LNPs, observed the internalization pathways used and tracked the particles as they co-localized with fluorescent protein tags on compartment-specific proteins, during endosomal sorting to the lysosome. The data revealed significant differences in the accumulation in subcellular compartments among the three formulations, which help to explain the observed effects LNP composition exerts on in vitro delivery efficiency.
Databáze: OpenAIRE
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