Search Institute (BIOMED), Hasselt University, Diepenbeek, BelgiumPF03.Extracellular vesicles derived from aged mesenchymal stem cells boost the regeneration capacity of mesenchymal stem cells Xiaoqin Wang; Chrysoula Tsirigoti; Forugh Vazirisani; Peter Thomsen; Karin Ekstr Division of Biomaterials, Institute of Clinical Sciences, Sahlgrenska Academy, University of Gothenburg, Gothenburg, SwedenBackground: Mesenchymal stem cells (MSCs) secret extracellular vesicles (EVs) which contribute towards the repair of several tissues. Studies have shown that in vitro ageing (passage variety of cells in culture) altered the characteristics of MSCs such as reduced proliferation and differentiation capacities. However, it is actually not however known if ageing impacts the secretion as well as the biological effects of MSC-derived EVs. Approaches: Conditioned media have been collected from three days serum no cost culture of human adipose-derived MSCs at P5 and P6 (low Protein tyrosine phosphatases Proteins manufacturer passages, LP), and P15 and P16 (high passages, HP). EVs were isolated by Exospin isolation kit and characterized by western blot and nanoparticle tracking evaluation. MSCs have been treated with each EVs_LP and EVs_HP with two various doses for six days as well as the proliferation capacity was evaluated by Cell Counting kit eight. Furthermore, the impact of EVs on osteogenic differentiation capacity was investigated by ALP assay just after 2 weeks of EVs remedy. Outcomes: Both MSC_LP and MSC_HP secreted EVs that had been good for CD63 and Flotillin 1, and unfavorable for Grp94. Particle quantification showed that MSC_HP secreted much more EVs than MSC_LP. Both EVs_LP and EVs_HP promoted MSC proliferation in comparison with nontreated group. In the low-dose remedy, EVs_LP and EVs_HPBackground: Tooth loss remains a major health problem given that present therapies cannot regenerate broken dental tissues like pulp and enamel. Thriving pulp regeneration is dependent upon angiogenesis, that is important for oxygen and nutrient provide. Proangiogenic characteristics have already been assigned to mesenchymal stem cells (MSCs) inside the dental pulp. So far, paracrine components, such as VEGF, happen to be identified as responsible angiogenic mediators. Having said that, a lot more current studies indicate that extracellular vesicles (EVs) created by bone marrow-derived MSCs (BMMSCs) also have the possible to induce neovascularisation. Therefore, we compared the angiogenic properties of EVs from dental pulp stem cells (DPSCs) with those of BMMSCs. Procedures: EVs had been isolated from serum-free conditioned medium of DPSCs and BMMSCs right after 48 h by differential ultracentrifugation. EV size and concentration were measured by nanoparticle tracking evaluation (NTA) and purity was confirmed by western blot with enrichment of classical EV markers CD9, CD63, CD81 and TSG101 and absence of non-EV marker mitochondrial complicated V. The functional effect of EVs on the migration of human umbilical vein endothelial cells (HUVECs), as a essential step in angiogenesis, was studied in a transwell technique. Results: Preliminary data recommend that EVs from DPSCs induce HUVEC migration (n = four). Having said that, this effect was much less compared to BMMSC EVs (n = two), which might be brought on by the decrease EV yield from DPSCs as measured by NTA. Uptake of DPSC EVs by Complement Factor H Related 1 Proteins Biological Activity HUVECs was confirmed with confocal microscopy. Summary/Conclusion: Our preliminary data show promising in vitro proangiogenic effects of DPSC EVs. In the future, we are going to evaluate the angiogenic components present in DPSC and BMMSC EVs and analyse their prospective to induce blood vessel gr.