Otoxic drug gentamicin and hypoxia, induce inflammatory response in SL pericytes. We then investigated the difference in exosome proteomics in normoxia and hypoxia. Strategies: SL pericytes have been obtained from Immortomouse Cells have been initially incubated with gentamicin in normoxic and hypoxic conditions as well as the degree of inflammatory response was assayed. FBS exosome depleted conditioned media was used for growing SL pericyte cultures in normoxic and hypoxic situations. Exosomes have been isolated from conditioned medium with basic differential ultracentrifugation and also the morphology examined by electron microscope. Exosome proteome was obtained with a LTQ Orbitrap Velos Pro ion-trap mass spectrometer. Final results: SL pericytes showed optimistic signal for the validated brain pericytes marker CD13. Incubation with all the ototoxic drug gentamicin induced SL phagocytic activity and improved the expression of cytokines for instance IL-6 as well as the LIF and MIP-2 and VEGF. The variations inside the exosome proteome from normoxic, hypoxic and gentamicin challenged cells was analysed with bioinformatics tools for identifying and visualising enriched GO terms and the proteins function on the exosome proteome. Summary: Exosome proteomes from normoxic hypoxic and gentamicin challenged SL pericites had been investigated in physiological situation and in inner ear pathological circumstances induced by hypoxia and ototoxic drug.Friday, May well 19,Poster Session F08 Intercellular and Inter-Organismal Crosstalk Chairs: Patricia Xander and Agnieszka Bronisz five:15:30 p.m.PF08.Human seminal plasma exosomes carry key proteins for PKCĪ¹ Compound spermatozoa capacitation Valentina Murdica1, Elisa Giacomini2, Alessandra Alteri2, Natasa Zarovni3, Andrea Salonia1, Paola Vigan and Riccardo Vago1 Urological Analysis Institute, IRCCS San Raffaele Hospital, Milano, Italy; Reproductive Sciences Laboratory, Division of Genetics and Cell Biology, IRCCS San Raffaele Hospital, Milano, Italy; 3Exosomics Siena SpA2Introduction: Male factor infertility is partially or fully accountable for as much as half of infertility situations and, amongst them, a relevant share final results from impairment of sperm maturation. In spite of assisted reproductive technologies being increasingly applied to cope with infertility, to date the fertilisation price is only partially successful by employing apparently regular semen. Recently, a certain interest has been directed for the role of exosomes in spermatozoa maturation and in conferring all round fertilisation capacity by the transfer of important molecules along the male reproductive tract Procedures: We collectedseminal plasma from normozoospermic sufferers upon getting approval by the neighborhood ethical committee and informed consent by the patient. Exosomes had been isolated and Fat Mass and Obesity-associated Protein (FTO) Formulation characterised by nanoparticle tracking evaluation, transmission electron microscopy and western blot. The uptake of exosomes derived from seminal plasma and labelled using a fluorescent dye by spermatozoa was monitored by immunofluorescence. Outcomes: Seminal plasma include both microvesicles and exosomes displaying canonical protein markers like CD9, CD63, Alix and TSG101. Additionally, exosomes, which represent a discrete population, carry proteins involved within the spermatozoa maturation and fertilisation capacity and inside the mechanism of anti-oxidative protection. Immediately after ejaculation, sperm cells are still receptive and can continue to obtain vesicle-delivered cargos. Certainly, we demonstrated that spermatozoa uptake exosomes derived from diverse sources. Conclu.