P –596 Association of oleic acid production in cumulus-granulosa cells with glutathione of in vitro matured oocytes

AbstractStudy questionDoes oleic acid production in cumulus-granulosa cells affect glutathione levels of in vitro matured oocytes?Summary answerOleic acid availability in cumulus-granulosa cells is associated with a higher glutathione level in in vitro matured oocytes.What is known alreadyThe monounsaturated fatty acid oleic acid is de novo synthesized by desaturation of stearic acid and can promote steroidogenesis and oocyte development in vitro. The endogenous antioxidant glutathione content in metaphase II oocyte is significantly higher than immature stages and is related to the normal oocyte maturation.Study design, size, durationMouse germinal vesicles were co-cultured for 24  hours, during in vitro maturation, with granulosa cells treated with a specific inhibitor of oleic acid synthesis alone or in combination with oleic acid.Participants/materials, setting, methodsFluorescence staining was used to assess the glutathione content of mouse metaphase II oocytes following in vitro maturation as an indicator of cytoplasmic maturation. Glutathione was stained using Cell Tracker Blue –CMAC for 30 min at 37 °C. After being washed in fresh media, stained oocytes were photographed by a fluorescence microscope. Cell area and associated fluorescence were quantified in 20 metaphase II mouse oocytes randomly chosen from in vitro matured oocytes for each condition.Main results and the role of chanceThe intracellular glutathione content was profoundly lower in metaphase II oo...
Source: Human Reproduction - Category: Reproduction Medicine Source Type: research