G of metabolomics and gene expression information are able to determine metabolic pathways that help to clarify the metabolic phenotype of PCa, and deliver novel therapeutics targets. This function also supports the study of urinary EVs as surrogate metabolic non-invasive markers for PCa tissue metabolism.PT05.Optimization of storage situations for extracellular vesicles Michel Bremer1; Verena B ger1; AndrG gens2; Samir El-Andaloussi2; Bernd Giebel1 Institute for Transfusion Medicine, University Hospital Essen, University of Duisburg-Essen, Essen, Germany; 2Clinical Research Center, Department for Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden, H sov en, Sweden; 3Institute for Transfusion Medicine, University Hospital Essen, Essen, GermanyBackground: EVs transmit particular information from their cells of origin to certain target cells and are key aspects inside a novel type of intercellular communication. Often, the EVs functional properties are analysed following their purification and normally they have been frozen and thaw prior to analysis. Storage, specially freezing BRaf Inhibitor supplier andthawing, might critically influence the integrity and functionality of respective EVs. Certainly, preliminary information of our group showed that long-time storage can decrease the number of particles recovered immediately after freezing and thawing. To optimize EV storage situations, we’ve studied the impact of various buffers and storing containers on recovery rates of stored eGFP-labelled EVs. Methods: In detail, eGFP-labelled EVs were harvested from supernatants of THP-1 cells, which had been transduced with CD63-eGFP encoding lentiviral particles. By ultracentrifugation purified EVs have been resuspended in six different buffers and aliquots of resulting suspensions transferred into 12 various plastic/glass containers each and every. Containers were either stored at 4 , -20 or -80 . Following a defined storage time and right after numerous freezing and thawing cycles, stored samples were analysed by nanoparticle tracking analysis (NTA). Final results: We observed a reduced recovery of particles for the duration of most storage circumstances. Independent from the storage temperatures, isotonic and pH-controlled buffers appeared preferable. Remarkably, the choice of storage containers as well as the storage temperature had massive effects on the particle concentrations and average size distributions of stored EVs. Even though EV rates were rather continual when stored at -80 , EV numbers varied drastically when stored at -20 or four , respectively. At present, we test for the functionality of stored EVs. Summary/conclusion: Combinations of storage containers, buffers and temperature drastically impact recovery prices of stored EVs. Funding: This study was funded by European Regional Improvement Fund 2014-2020 (EFRE) and European Union.Thursday, 03 MayPT06: EVs in Cellular Differentation and Organ Development Chairs: Ana Gamez Valero; Guillaume van Niel Location: CD40 Inhibitor Synonyms Exhibit Hall 7:158:PT06.Driving patched to the bottom: vesicular trafficking to polarize Hh reception Ana C. Gradilla; Laura Gonz ez-M dez; Isabel Guerrero Centro de Biologia Molecular Severo Ochoa (CSIC-UAM), Madrid, SpainBackground: The Hedgehog (Hh) signalling pathway is essential for early animal development and tissue maintenance within the adult. The lipid-modified Hh acts as a morphogen and signals within a graded manner, reaching differential responses inside the getting cell in line with ligand concentration. As a result, the extracellular ligand distribution from the membrane anchored Hh.