Isolation of extracellular vesicles from citrus limon fruit through membrane-based techniques.

The project is aimed at the development of a membrane-based protocol for the isolation of plant-derived extracellular vesicles. In particular, citrus limon fruit is selected as source material, due to its beneficial phyto-properties, availability and cheapness. In the chosen strategy the focus is placed upon the development of a protocol potentially suitable for large-scale operations, conversely to differential ultracentrifugation method, the benchmark in extracellular vesicles production, whose applicability is limited to lab-scale operations. In this context dead-end and crossflow MF/UF units are compared, both as concentration and purification steps. Followingly, the isolated vesicles ware analysed and characterized with SEC liquid chromatography and both physical and biochemical methods. Finally, passive cargo loading techniques are applied to the isolated vesicles to test their encapsulation capability with bioactive phytocomponents, as curcumin

Large-scale production of extracellular vesicles: Report on the “massivEVs” ISEV workshop

Extracellular vesicles (EVs) large-scale production is a crucial point for the translation of EVs from discovery to application of EV-based products. In October 2021, the International Society for Extracellular Vesicles (ISEV), along with support by the FET-OPEN projects, “The Extracellular Vesicle Foundry” (evFOUNDRY) and “Extracellular vesicles from a natural source for tailor-made nanomaterials” (VES4US), organized a workshop entitled “massivEVs” to discuss the potential challenges for translation of EV-based products. This report gives an overview of the topics discussed during “massivEVs”, the most important points raised, and the points of consensus reached after discussion among academia and industry representatives. Overall, the review of the existing EV manufacturing, upscaling challenges and directions for their resolution highlighted in the workshop painted an optimistic future for the expanding EV field