An efficient method to isolate lemon derived extracellular vesicles for gastric cancer therapy

Plant-derived extracellular vesicles (PDEVs) have great potential for clinical applications. Ultracentrifugation, considered the gold standard method for the preparation of PDEVs, is efficacious but time-consuming and highly instrument-dependent. Thus, a rapid and handy method is needed to facilitate the basic researches and clinical applications of PDEVs. In this study, we combined electrophoretic technique with 300 kDa cut-off dialysis bag (named ELD) for the isolation of PDEVs, which was time-saving and needed no special equipment. Using ELD, lemon derived extracellular vesicles (LDEVs) could be isolated from lemon juice. Nanoparticle tracking analysis and transmission electron microscopy confirmed that the method separated intact vesicles with a similar size and number to the standard method-ultracentrifugation. LDEVs caused the gastric cancer cell cycle S-phase arrest and induced cell apoptosis. The anticancer activities of LDEVs on gastric cancer cells were mediated by the generation of reactive oxygen species. In addition, LDEVs were safe and could be remained in gastrointestinal organs. ELD was an efficient method for the isolation of LDEVs, and could be carried out in any routine biological laboratory as no special equipment needed. LDEVs exerted anticancer activities on gastric cancer, indicating the great potentials for clinical application as edible chemotherapeutics delivery vehicle.