Rresed Pontificia PIM2 web Universidad Cat ica de Chile; University Medical Center of Groningen, Groningen, Netherlands; bUMCG, Groningen, Netherlands; Pontificia Universidad Cat ica de Chile/Universidad Bernardo O iggins, SANTIAGO, Chile; dPontificia Universidad Cat ica de Chile, Santiago, Chile; eUniversity Health-related Center Groningen, Groningen, Netherlandsc aPS01.Human telomerized cells for production of extracellular vesicles Regina Grillaria, Susanne Neubertb, Matthias Wiesera and Johannes GrillaribaEvercyte GmbH, Vienna, Austria; bChristian Doppler Laboratory on Biotechnology of Skin Aging, University of Organic Sources and Life Sciences, Vienna (BOKU), Vienna, AustriaIntroduction: Human cells are of ever escalating importance as in vitro test system to represent the in vivo predicament. Moreover, extremely differentiated cells are also crucial production systems for αvβ5 drug complex biopharmaceuticals. Even so, the use of such cell systems are limited because of the truth that the cells enter replicative life span and as a result can only be propagated to get a restricted number of population doublings in vitro, which restricted standardization of experiments also as production processes. Additionally, reports have shown that the number of secreted vesicles drastically lowered with escalating age of regular cells.Introduction: Background: Transition from isolated steatosis (IS) to non-alcoholic steatohepatitis (NASH) is often a crucial concern in non-alcoholic fatty liver illness (NAFLD). Current observations in individuals with obstructive sleep apnea syndrome (OSAS), recommend that hypoxia might contribute to disease progression mostly by way of activation of hypoxia inducible factor 1 (HIF-1)-related pathways. Release of extracellular vesicles (EV) by injured hepatocytes may well be involved in NAFLD progression. Aim: To explore no matter if hypoxia modulates the release of EV from totally free fatty acid (FFA)-exposed hepatocytes and assess cellular crosstalk among hepatocytes and LX-2 cells (human hepatic stellate cell line). Procedures: HepG2 cells had been treated with FFAs (250 M palmitic acid + 500 M oleic acid) and chemical hypoxia (CH) was induced with Cobalt (II) Chloride, that is an inducer of HIF-1. Induction of CH was confirmed by Western blot (WB) of HIF-1. EV isolation and quantification was performed by ultracentrifugation and nanoparticle tracking analysis respectively. EV characterization was performed by electron microscopy and WB of CD-81 marker. LX-2 cells have been treated with 15 g/ml of EV from hepatocytes obtained from unique groups and markers of pro-fibrogenic signalling were determined by quantitative PCR (qPCR), WB and immunofluorescence (IF). Benefits: FFA and CH-treatment of HepG2 cells increased gene expression of IL-1 and TGF-1 inJOURNAL OF EXTRACELLULAR VESICLESHepG2 cells and increased the release of EV compared to non-treated HepG2 cells. Treatment of LX-2 cells with EV from FFA-treated hypoxic HepG2 cells improved gene expression of TGF-1, CTGF, -SMA and Collagen1A1 in comparison with LX-2 cells treated with EV from non-treated hepatocytes or LX-2 cells exposed to EV-free supernatant from FFA-treated hypoxic HepG2 cells. Moreover, EV from FFA-treated hypoxic HepG2 cells increased Collagen1A1 and -SMA protein levels.Summary/conclusion: CH promotes EV release from HepG2 cells. EV from hypoxic FFA-treated HepG2 cells evoke pro-fibrotic responses in LX-2 cells. Further genomic and proteomic characterization of EV released by steatotic cells beneath hypoxia are necessary to additional.