Te srl, Turin, Italyb aIntroduction: Extracellular vesicles (EVs) are particles released by cells that carry a complicated cargo of molecules and mediate intercellular communication. Lately, they’ve raised good interest as drug delivery systems and a number of engineering approaches are currently beneath investigation. Various factors, nonetheless, influence the transfection yield, which includes protocol variability and EV damage. Solutions: The electroporation was investigated as method to straight load miRNAs in plasma-derived EVs. Distinct parameters (voltage and quantity of pulses) were compared for their impact on EV morphology and loading capacity of a synthetic miRNA, cel-39, including miRNA enrichment in EVs and its transfer to target cells. Next, analyses were performed to evaluated the transfection effect on EV endogenous cargo along with the exogenous miRNA protection from RNAse degradation. Then, EVs have been loaded with antitumour miRNAs and their proapoptotic impact was evaluated on a cell line of hepatocellular carcinoma, HepG2 cells.JOURNAL OF EXTRACELLULAR VESICLESResults: The comparison of distinctive electroporation settings demonstrated the value of picking the far more proper protocol parameters to acquire an effective EV transfection yield, understood as both molecules loading and EV harm. In unique, we observed the superiority of one particular electroporation protocol (working with 750 Volt and 10 pulses) that permitted probably the most efficient miRNA packaging and transfer to target cells, devoid of structurally damaging EVs. One of the most efficient electroporation protocol was also verified to let a more efficient miRNA loading in respect to incubation, superior safeguarding miRNA from enzymatic digestion. Furthermore, our findings suggested that electroporation preserved the na e EV cargo, like RNAs and proteins, and did not alter their uptake in cells. EVs engineered with antitumor miRNAs (miR-31 and miR-451a) successfully promoted the apoptosis of HepG2 cells, downregulating their target genes connected to apoptotic pathways. Summary/Conclusion: In conclusion, our findings indicate an efficient and functional miRNA encapsulation in plasma-derived EVs following an electroporation protocol that preserves EV integrity. Funding: Associazione Italiana per la Ricerca sul Cancro (A.I.R.C.), Unicyte AG (Switzerland)PS01.LAT1/CD98 Proteins supplier Improvement of a platform for exosome engineering making use of a novel and selective scaffold protein for surface show Kevin Dooley, Ke Xu, Sonya Haupt, Shelly Integrin beta 2/CD18 Proteins Recombinant Proteins Martin, Russell McConnell, Nuruddeen Lewis, Christine McCoy, Chang Ling Sia, Jorge Sanchez-Salazar, Nikki Ross, Rane Harrison, Bryan Choi, Damian Houde, John Kulman and Sriram Sathyanarayanan Codiak BioSciences, Cambridge, USAfragments thereof were expressed within a cell line plus the minimum PrX domain needs for exosomal enrichment were determined. Leveraging PrX as a scaffold for exosome surface show, we developed our engEx platform to create engineered exosomes functionalized with a selection of pharmacologic payloads including enzymes, antibodies, sort I cytokines and TNF superfamily members. Biological activity of those engineered exosomes was assessed in an array of in vitro assays and compared to previously described scaffolds. Results: Stable expression of PrX in an exosome creating cell line resulted in 200-fold enrichment of PrX on secreted exosomes. Interestingly, overexpression of PrX structural paralogs didn’t lead to comparable levels of enrichment, suggesting PrX is special. Exos.