Uptake of Liposomally Entrapped Fluorescent Antisense Oligonucleotides in NG108-15 Cells: Conventional versus pH-Sensitive.

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Antisense oligodeoxynucleotides (asODN) are novel therapeutic agents designed to alter RNA metabolism, ultimately resulting in decreased production of disease-associated gene products. To investigate internalisation of liposomally delivered asODN in NG108-15 cells, a hybrid cell line of mouse neuroblastoma and rat glioma, and assure that uptake of marker corresponds to that of antisense, we compared the cellular uptake of fluorescently labelled marker (fluorescein isothiocyanate (FITC)–dextran) and antisense oligonucleotide (FITC–asODN), entrapped either in conventional soy phosphatidylcholine (SPC) liposomes or pH-sensitive liposomes (composed of dioleoylphosphatidylethanolamine and cholesteryl hemisuccinate in a molar ratio of 3 : 2). Both SPC and pH-sensitive liposomes were prepared by a modified freeze-thawing method. Entrapment efficiencies (about 20% of the original material) did not depend on the liposome compositions and fluorescent material used. Fluorescence activated cell sorting (FACS) analysis was used to quantify the association of fluorescent material with the NG108-15 cells, whereas confocal microscopy gave insight on the location of cell associated-fluorescence. Conventional liposomes failed to deliver fluorescent material into the cells, but in contrast, pH-sensitive liposomes significantly improved the uptake of both FITC–dextran and FITC–asODN, with the uptake of liposomal FITC–dextran being greater than the uptake of liposomal FITC–asODN. These results suggest that pH-sensitive liposomes can be applied as a carrier system in the delivery of genetic material into the cells.
公益社団法人日本薬学会の論文

Uptake of Liposomally Entrapped Fluorescent Antisense Oligonucleotides in NG108-15 Cells: Conventional versus pH-Sensitive.

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