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 6, 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: Around the road towards the usage of extracellular vesicles (EVs) for regenerative medicine, technological hurdles remain unsolved: high-yield, higher purity and cost-effective mGluR2 Biological Activity production of EVs. Solutions: Pursuing the analogy with shear-stress induced EV release in blood, we’re developing a mechanical-stress EV triggering cell culture approach in scalable and GMP-compliant bioreactors for costeffective and higher yield EV production. The third generation setup allows the production of up to 300,000 EVs per Mesenchymal Stem Cell, a 100-fold boost in comparison to classical approaches, i.e physiological spontaneous release in depleted media (about 2000 EVs/ cell), with a high purity ratio 1 10e10 p/ Outcomes: We investigated in vitro the regenerative possible of high yield mechanically induced MSC-EVs by demonstrating an equal or improved efficiency in comparison to classical EVs with all the exact same quantity of EVs. The regenerative properties of mechanically induced MSCEVs was confirmed in vivo within a murine model of chronic heart failure demonstrating that higher, medium shear pressure EVs and serum starvation EVs or mMSCs had exactly the same effect utilizing local injection. We later on tested the effect of your injection route plus the use of xenogenic hMSC-EVs on their efficiency inside the identical model of murine chronic heart failure. Heart functional parameters were analysed by ultrasound two months (1 month post EV injection) post infarction. Interestingly, hMSCEVs had the same effect compared to mMSC-EVs in local injection, displaying that xeno-EVs in immunocompetent mices was well tolerated. In addition, hMSC EV IV injection was as efficient as nearby intra-myocardium κ Opioid Receptor/KOR Purity & Documentation muscle injection with an increase within the left ventricular ejection fraction of 26 in comparison to pre-treatment values, whereas PBS injected controls lost 13 . Summary/Conclusion: We demonstrated an equal or superior regenerative effect of high yield mechanically made EVs compared to spontaneously released EVs or parental cells in vitro and in vivo, and great tolerance and efficacy of hMSC EV each with neighborhood and IV injection. This distinctive technologies for EV production combines decisive assets for clinical translation of EV-based regenerative medicine : a GMP-compliant setup, higher density cell culture, high yield re.
