Rresed Pontificia Universidad Cat ica de Chile; University Health-related Center of Groningen, Groningen, Netherlands; bUMCG, Groningen, Netherlands; Pontificia Universidad Cat ica de Chile/Universidad FSH Receptor Proteins Gene ID 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 rising significance as in vitro test program to represent the in vivo circumstance. In addition, extremely differentiated cells are also crucial production systems for complicated biopharmaceuticals. Having said that, the usage of such cell systems are restricted due to the reality that the cells enter replicative life span and as a result can only be propagated for a restricted quantity of population doublings in vitro, which limited standardization of experiments too as production processes. Additionally, reports have shown that the number of secreted vesicles considerably lowered with rising age of typical cells.Introduction: Background: Transition from isolated steatosis (IS) to non-alcoholic steatohepatitis (NASH) is actually a essential problem in non-alcoholic fatty liver disease (NAFLD). Current observations in individuals with obstructive sleep apnea syndrome (OSAS), recommend that hypoxia may possibly contribute to illness progression mainly by means of activation of hypoxia inducible issue 1 (HIF-1)-related pathways. Release of extracellular vesicles (EV) by injured hepatocytes may well be involved in NAFLD progression. Aim: To explore whether hypoxia modulates the release of EV from cost-free fatty acid (FFA)-exposed hepatocytes and assess cellular crosstalk involving hepatocytes and LX-2 cells (human hepatic stellate cell line). Strategies: HepG2 cells were treated with FFAs (250 M palmitic acid + 500 M oleic acid) and chemical hypoxia (CH) was induced with Cobalt (II) Chloride, which is an inducer of HIF-1. Induction of CH was confirmed by IgG2C Proteins Recombinant Proteins 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 different groups and markers of pro-fibrogenic signalling had been determined by quantitative PCR (qPCR), WB and immunofluorescence (IF). Results: FFA and CH-treatment of HepG2 cells elevated gene expression of IL-1 and TGF-1 inJOURNAL OF EXTRACELLULAR VESICLESHepG2 cells and enhanced the release of EV in comparison with non-treated HepG2 cells. Treatment of LX-2 cells with EV from FFA-treated hypoxic HepG2 cells elevated gene expression of TGF-1, CTGF, -SMA and Collagen1A1 when compared 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. Furthermore, EV from FFA-treated hypoxic HepG2 cells elevated 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. Additional genomic and proteomic characterization of EV released by steatotic cells below hypoxia are necessary to further.