Stability of Plant Leaf-Derived Extracellular Vesicles According to Preservative and Storage Temperature

Plant-derived extracellular vesicles (EVs) are capable of efficiency delivering mRNAs, miRNAs, bioactive lipids, and proteins to mammalian cells. Plant-derived EVs critically contribute to the ability of plants to defend against pathogen attacks at the plant cell surface. They also represent a novel candidate natural substance that shows potential to be developed for food, cosmetic, and pharmaceutical products. However, although plant-derived EVs are acknowledged as having potential for various industrial applications, little is known about how their stability is affected by storage conditions. In this study, we evaluated the stability of Dendropanax morbifera leaf-derived extracellular vesicles (LEVs) alone or combined with the preservatives, 1,3-butylene glycol (to yield LEVs-1,3-BG) or TMO (LEVs-TMO). We stored these formulations at -20, 4, 25, and 45 degrees C for up to 4 weeks, and compared the stability of fresh and stored LEVs. We also assessed the effect of freeze-thawing cycles on the quantity and morphology of the LEVs. We found that different storage temperatures and number of freeze-thawing cycles altered the stability, size distribution, protein content, surface charge, and cellular uptake of LEVs compared to those of freshly isolated LEVs. LEVs-TMO showed higher stability when stored at 4 degrees C, compared to LEVs and LEVs-1,3-BG. Our study provides comprehensive information on how storage conditions affect LEVs and suggests that the potential industrial applications of plant-derived EVs may be broadened by the use of preservatives.

Automated summary

Plant-derived extracellular vesicles (EVs) have the ability to efficiently deliver various substances to mammalian cells and play a critical role in plant defense against pathogens. They show potential for industrial applications in food, cosmetic, and pharmaceutical products. However, little is known about the stability of plant-derived EVs under different storage conditions. This study evaluated the stability of Dendropanax morbifera leaf-derived extracellular vesicles (LEVs) and their formulations with preservatives under different storage temperatures. The results provide comprehensive information on the stability of LEVs and suggest the use of preservatives to broaden the potential industrial applications of plant-derived EVs.