RRC ID 81785
Author Okami K, Fumoto S, Yamashita M, Nakashima M, Miyamoto H, Kawakami S, Nishida K.
Title One-Step Formation Method of Plasmid DNA-Loaded, Extracellular Vesicles-Mimicking Lipid Nanoparticles Based on Nucleic Acids Dilution-Induced Assembly.
Journal Cells
Abstract We propose a nucleic acids dilution-induced assembly (NADIA) method for the preparation of lipid nanoparticles. In the conventional method, water-soluble polymers such as nucleic acids and proteins are mixed in the aqueous phase. In contrast, the NADIA method, in which self-assembly is triggered upon dilution, requires dispersion in an alcohol phase without precipitation. We then investigated several alcohols and discovered that propylene glycol combined with sodium chloride enabled the dispersion of plasmid DNA and protamine sulfate in the alcohol phase. The streamlined characteristics of the NADIA method enable the preparation of extracellular vesicles-mimicking lipid nanoparticles (ELNPs). Among the mixing methods using a micropipette, a syringe pump, and a microfluidic device, the lattermost was the best for decreasing batch-to-batch differences in size, polydispersity index, and transfection efficiency in HepG2 cells. Although ELNPs possessed negative ζ-potentials and did not have surface antigens, their transfection efficiency was comparable to that of cationic lipoplexes. We observed that lipid raft-mediated endocytosis and macropinocytosis contributed to the transfection of ELNPs. Our strategy may overcome the hurdles linked to supply and quality owing to the low abundance and heterogeneity in cell-based extracellular vesicles production, making it a reliable and scalable method for the pharmaceutical manufacture of such complex formulations.
Volume 13(14)
Published 2024-7-11
DOI 10.3390/cells13141183
PII cells13141183
PMID 39056764
PMC PMC11274598
MeSH DNA* / chemistry DNA* / metabolism Extracellular Vesicles* / metabolism Hep G2 Cells Humans Lipids* / chemistry Liposomes Nanoparticles* / chemistry Nucleic Acids / chemistry Nucleic Acids / metabolism Plasmids* / genetics Transfection* / methods
IF 4.366
Resource
Human and Animal Cells Hep G2(RCB1886)