Otentially damaging plasmid DNA and off-target toxicity. The findings move this method closer to clinical transfer. Funding: NIH NCATS UH3TR000902.OF11.Higher yield hMSC derived mechanically induced xenografted extracellular vesicles are well tolerated and induce potent regenerative impact in vivo in neighborhood or IV p38α supplier injection within 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 remain unsolved: high-yield, high purity and cost-effective production of EVs. Strategies: 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 improve in comparison to classical strategies, i.e physiological spontaneous release in depleted media (around 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 elevated efficiency in comparison with classical EVs using the exact same level of EVs. The regenerative properties of mechanically induced MSCEVs was confirmed in vivo P2X1 Receptor MedChemExpress inside a murine model of chronic heart failure demonstrating that high, medium shear anxiety EVs and serum starvation EVs or mMSCs had the exact same impact making use of neighborhood injection. We later on tested the impact in the injection route plus the use of xenogenic hMSC-EVs on their efficiency within the similar 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 exactly the same effect in comparison to mMSC-EVs in neighborhood injection, showing that xeno-EVs in immunocompetent mices was properly tolerated. Furthermore, hMSC EV IV injection was as effective as regional intra-myocardium muscle injection with a rise inside 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 impact of higher yield mechanically made EVs when compared with spontaneously released EVs or parental cells in vitro and in vivo, and excellent tolerance and efficacy of hMSC EV both with regional and IV injection. This one of a kind technologies for EV production combines decisive assets for clinical translation of EV-based regenerative medicine : a GMP-compliant setup, higher density cell culture, higher yield re.
