Fluorescence studies on the mechanism of liposome-cell interactions in vitro.
Sonicated unilamellar liposomes containing fluorescent lipid analogs or biotinyl phosphatidylethanolamine as a ligand for fluorescein avidin have been used to study the mechanism of interaction of phospholipid vesicles with eucaryotic cells. Microscopy revealed that after short incubations the fluorescence was associated with the cell surface in a punctate as opposed to a uniform staining pattern. Fluid vesicles, regardless of charge, were found to associate with cells to the same degree. Solid neutral and negatively charged vesicles associated to a 3-fold greater extent, while solid positively charged vesicles associated to a 10-fold greater extent than fluid vesicles. Fluorescence recovery after photobleaching, a technique used to measure the lateral mobility of cell surface components, was used to measure the lateral mobility of the associated fluorescence probes. No recovery was observed, implying that greater than 90% of the fluorescent lipid analogs are not free to diffuse over distances of the order of 1 micrometer. When these analogs were introduced into the cell membrane by an ethanol-injection technique, rapid and full recovery after photobleaching was observed. This can be accounted for by a lateral diffusion coefficient characteristic of phospholipids in model and biomembranes. The image and photobleaching results suggest that the majority of liposomes that become cell-associated under the conditions used here are adsorbed on the surface. The consequences of this binding for liposome-mediated delivery of molecules into the cytoplasm or plasma membrane of the cell are discussed.
D000818:Animals; D001710:Biotin; D002232:Carbocyanines; D002462:Cell Membrane / Q000378:metabolism*; D005456:Fluorescent Dyes; D007739:L Cells / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D010169:Palmitic Acids / Q000378:metabolism; D010713:Phosphatidylcholines / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000378:metabolism; D013229:Stearic Acids / Q000378:metabolism
D002232:Carbocyanines; D005456:Fluorescent Dyes; D008081:Liposomes; D010169:Palmitic Acids; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D013229:Stearic Acids; D001710:Biotin