Otentially damaging plasmid DNA and off-target toxicity. The findings move this strategy closer to clinical transfer. Funding: NIH NCATS UH3TR000902.OF11.High yield hMSC derived mechanically induced xenografted extracellular vesicles are nicely tolerated and induce potent regenerative impact in vivo in local or IV injection inside a model of chronic heart failure Max Piffouxa, Iris Marangonb, Nathalie Mougenotc, Claire Wilhelmd, Florence Gazeaue, Onnik Agbulutf and Amanda Brun-Silvaga Laboratoire Mati e et Syst es Complexes, CNRS UMR 7047 UniversitParis Diderot, Paris, France; bUniversitSorbonne Paris Cit Laboratoire Mati e et Syst es Complexes, CNRS UMR 7047 UniversitParis Diderot, France; cSorbonne Universit , UniversitPierre et Marie Curie Paris six, Plateforme PECMV, UMS28, Paris, France; dlaboratoire Mati e et Syst es Complexes, paris, France; eUniversitSorbonne Paris Cit Laboratoire Mati e et Syst es Complexes, CNRS UMR 7047 UniversitParis Diderot, Paris, France; fUniversitSorbonne Paris Cit Laboratoire Mati e et Syst es Complexes, CNRS UMR 7047 UniversitParis Diderot, Paris, France; 7UniversitSorbonne Paris Cit Laboratoire Mati e et Syst es Complexes, CNRS UMR 7047 UniversitParis Diderot, Paris, FranceIntroduction: On the road towards the use of extracellular vesicles (EVs) for regenerative medicine, technological hurdles stay unsolved: high-yield, higher purity and cost-effective production of EVs. Solutions: Pursuing the analogy with shear-stress induced EV release in blood, we’re building a mechanical-stress EV triggering cell culture strategy in scalable and GMP-compliant bioreactors for costeffective and higher yield EV production. The third generation setup enables the production of as much as 300,000 EVs per Mesenchymal Stem Cell, a 100-fold enhance compared to classical solutions, i.e physiological spontaneous release in CD29/Integrin beta-1 Proteins Species depleted media (about 2000 EVs/ cell), using a high purity ratio 1 10e10 p/ Benefits: We investigated in vitro the regenerative possible of high yield mechanically induced MSC-EVs by demonstrating an equal or increased efficiency when compared with classical EVs with the same volume of EVs. The regenerative properties of mechanically induced MSCEVs was confirmed in vivo in a murine model of chronic heart failure demonstrating that high, medium shear strain EVs and serum starvation EVs or mMSCs had the exact same impact working with nearby injection. We later on tested the effect with the injection route and also the use of xenogenic hMSC-EVs on their efficiency within the same model of murine chronic heart failure. Heart functional parameters had been analysed by ultrasound two months (1 month post EV injection) post infarction. Interestingly, hMSCEVs had exactly the same impact when compared with mMSC-EVs in regional injection, showing that xeno-EVs in immunocompetent mices was well tolerated. In addition, hMSC EV IV injection was as efficient as nearby intra-myocardium muscle injection with a rise within the left ventricular ejection fraction of 26 in comparison with pre-treatment values, whereas PBS injected controls lost 13 . Summary/Conclusion: We demonstrated an equal or superior regenerative effect of high yield mechanically created EVs in comparison with spontaneously BST-2/CD317 Proteins Recombinant Proteins released EVs or parental cells in vitro and in vivo, and fantastic tolerance and efficacy of hMSC EV both with regional and IV injection. This special technologies for EV production combines decisive assets for clinical translation of EV-based regenerative medicine : a GMP-compliant setup, high density cell culture, high yield re.
