LAUSR

Fusion between giant unilamellar vesicles (GUVs) can incorporate and mix components of biochemical reactions. Recently, GUV fusion induced by freeze‐thawing (F/T) was employed to construct artificial cells that can easily and repeatedly fuse GUVs with efficient content mixing. However, GUVs were ruptured during F/T, and the inner contents leaked. Herein, we investigated the effects of charged lipids on GUV fusion via F/T. The presence of 10 %‐50 % (w/w%) negatively charged lipids in GUV membranes, mainly composed of the neutral charged lipid 1‐palmitoyl‐2‐oleoyl‐sn‐glycero‐3‐phosphocholine (POPC), improved resistance to GUV rupture and decreased inner content leakage. Furthermore, we found that the presence of positively charged lipids in GUV membranes elevated GUV rupture compared with F/T between GUVs containing POPC alone. Modified GUVs may better incorporate nutrients and lipid membranes with less damage following GUV fusion via F/T, providing an improved artificial model.