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Small extracellular vesicles (sEVs) are known to play an important role inthe communication between distant cells and to deliver biological informationthroughout the body. To date, many studies have focused on the role of sEVscharacteristics such as cell origin, surface composition, and molecular cargoon the resulting uptake by the recipient cell. Yet, a full understanding of thesEV fusion process with recipient cells and in particular the role of cellmembrane physical properties on the uptake are still lacking. Here we explorethis problem using sEVs from a cellular model of triple-negative breast cancerfusing to a range of synthetic planar lipid bilayers both with and withoutcholesterol, and designed to mimic the formation of raft-like nanodomains incell membranes. Using time-resolved Atomic Force Microscopy we were able totrack the sEVs interaction with the different model membranes, showing theprocess to be strongly dependent on the local membrane fluidity. The strongestinteraction and fusion is observed over the less fluid regions, with sEVs evenable to disrupt ordered domains at sufficiently high cholesterol concentration.Our findings suggest the biophysical characteristics of recipient cellmembranes to be crucial for sEVs uptake regulation.

Lipid bilayer fluidity and degree of order regulates small EVs adsorption on model cell membrane