O-233 Follicular fluid extracellular vesicles as a potential tool for human spermatozoa selection in vitro

Abstract Study question What characteristics and features of follicular fluid (FF) extracellular vesicles (EVs) women different ages can affect the functional spermatozoa? Summary answer The size, lipid composition, progesterone levels FF EVs change with a woman's age fertilizing ability spermatozoa. is known already Aging influences on molecular composition FF, reducing fertilization processes occurring in fallopian tubes. an important factor activating spermatozoa during oocyte fertilization, enhancing process sperm hyperactivation capacitation. one main chemoattractants female genital tract, attracting to starting processes. Our previous studies have shown significant effect young compared advanced maternal (AMA) motility binding acrosoma also was detected. design, duration were obtained by sequential centrifugation at rotational speeds. fraction isolated, washed differential density gradient, suspended saline concentration 10*6/ml incubated (1:2) 37 °C CO2-incubator for 1 h. Next steps: immunofluorescent staining, test acrosomal reaction after incubation, NTA analysis, profile measuring ages. Participants/materials, setting, methods All study participants signed voluntary informed consent use biological samples research purposes. Sperm isolated from seminal (n = 21) aged 28-36. 14) 22-30 AMA patients 42-47. used this work include immunofluorescence microscopy, nanoparticle tracking flow cytometry, liquid chromatography–mass spectrometry, statistical data analysis. Lipids extracted using modified Folch extraction protocol. Main results role chance Progesterone significantly higher controls (3951000 a.u., 598500 6701 a.u.). This may explain increase incubation EVs. difference size (144.78 nm ± 9.92 VS 127.03 17.17) women's more therapeutic A total 389 vesicle lipids identified. level cholesterol esters, cholesterol, lysophosphatidylcholines, phosphatidylcholines 1,5 times women. di- triglycerides, ceramides, oxidized forms spontaneous almost 3 lower control (11,21% 32,88%). supports hypothesis that restrain premature Age-related changes are reflected reproductive processes, which leads functions Limitations, reasons caution Small amount data. To be completely confident importance participation activation spermatozoa, it necessary perform proteomic analyses discuss possible mechanisms underlying effects groups characteristics. Wider implications findings Further help not only complement fundamental understanding impact morphofunctional characteristics, but develop tool improving ART outcomes couples male infertility. solutions could donors. Trial registration number applicable