The MBs were fabricated by dissolving the dipalmitoyl-phosphatidyl-choline (DPPC) (Avanti Polar Lipids, AL, USA), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(poly(ethyleneglycol))-2000] (DSPE-PEG2000) (Avanti Polar Lipids) and 1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP) (Avanti Polar Lipids) in chloroform at a molar ratio of 9:2:1, and then draining to generate a lipid film 30 (link). The film was then dissolved with 1 wt% glycerol-containing phosphate-buffered saline (PBS). Subsequently, the solution was degassed and refilled with perfluoropropane (C3F8). The MBs were formed by intense mechanical shaking using an agitator for 45 s. Finally, to separate from unreacted lipids, centrifugation was applied to the MBs at 6,000 rpm (2000 g) for 3 min.
Plasmid DNA (0-10000 ng) was gently mixed with 108 MBs for 30 min, and was centrifuged at 6,000 rpm (2000 g) for 1 min for separating unattached plasmid DNA. The normal MBs without plasmid DNA payload were prepared for comparison.
Plasmid DNA (0-10000 ng) was gently mixed with 108 MBs for 30 min, and was centrifuged at 6,000 rpm (2000 g) for 1 min for separating unattached plasmid DNA. The normal MBs without plasmid DNA payload were prepared for comparison.
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