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

Abstract

Background: 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. Results: In this study, we combined electrophoretic technique with 300 kDa cutoff 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 methodultracentrifugation. 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. Conclusions: 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.

Extracellular Vesicles from Plants: Current Knowledge and Open Questions

“…They centrifuged lemon juice at 3000× g for 10 min and 10,000× g for 20 min and filtered the supernatant through a 0.22 µm pore size filter. Then, the juice was placed in a 300 kDa dialysis bag placed in a gel holder cassette with a current of 300 mA [18].…”

“…Several studies have emphasized the anti-cancer properties of EVs derived from different plants, thus enabling them to become potential therapeutic compounds in combination with current treatments in cancer management [4,5,14,[17][18][19]39].…”

Plant-Derived Nano and Microvesicles for Human Health and Therapeutic Potential in Nanomedicine

“…In the last decade, different approaches, mostly based on ultracentrifugation techniques, schematized in Figure 1 , have been applied to isolate distinct classes of vesicles from tissues, organs, apoplastic fluid, and juices of several plant species such as ginger and carrot roots [ 2 ], grape [ 16 ] and citrus fruits [ 17 , 18 , 19 , 20 ], ginseng [ 21 ], apple [ 22 ], wheat [ 23 ] and many others. Very recently, round-shaped EVs were isolated by differential ultracentrifugation of a sampling solution containing root exudates of hydroponically grown tomato plants [ 24 ].…”

Extracellular Vesicles: “Stealth Transport Aircrafts” for Drugs

“…LDEVs caused s-phase arrest of gastric cancer cell cycle and induced cell apoptosis. LDEVs could be retained in the organs of the gastrointestinal tract and had strong anti-tumor activity against gastric cancer [82]. The isolated plant EVs can also be used after being engineered.…”