Rapid Access to Giant Unilamellar Liposomes with Upper Size Control: Membrane-Gated, Gel-Assisted Lipid Hydration

Combining gel-assisted lipid hydration with membrane-based lipid extrusion, we demonstrate here a general procedure for rapid preparation of giant unilamellar liposomes with upper size control. Featured in this procedure are planar lipid stacks deposited on poly­(vinyl alcohol) gel, which are further laminated atop with microporous polycarbonate membranes. Control of liposome size is thus realized through the uniform-sized pores of the latter, which provide the only access for the underlying lipids to enter the main aqueous phase upon hydration. Production of both single-phased and biphasic (Janus) liposomes using several commonly employed model lipids, including 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine (POPC) and cholesterol, is presented. The size distribution, yield and lamellarity of these liposome products are characterized and analyzed in detail by confocal fluorescence microscopy. This procedure thus offers a simple and fast alternative route to giant unilamellar liposomes with upper size control.