Combined space-organized system for animals oocytes maturation in vitro
Aim. The aim of the work was to create a spaсe-organized system for oocytes maturation based on rabbits granulosa cells, hyaluronic acid (for structural organization of this system) and insulin, as an important growth factor. Methods. Cultivation of granulosa cells and oocytes maturation in vitro, biochemical determination of lactate dehydrogenase activity and cholesterol concentration, and determination of progesterone and estradiol concentrations by enzyme immunoassay were done. Results. Hyaluronic acid in concentration of 0.5 μg/ml increased proliferation of granulosa cells and progesterone concentration in the conditioned medium on 72 h of cultivation, and estradiol on 48 and 72 hours of cultivation was found. The degree of oocyte maturation to metaphase-2 was 85% by the influence of 10 μg/ml insulin. Same concentration of insulin increased granulosa cells proliferation, estradiol, progesterone and cholesterol concentration in the conditioning medium after 24 hours of cultivation. Conclusions. The optimal parameters of the combined spaceorganized culture system are 1 million cells/ml of granulose with the addition of 0.5 μg/ml hyaluronic acid and 10 μg/ml insulin, which creates the optimal conditions for the maturation of oocytes to metaphase-2 cell cycle.
Keywords: oocytes, granulosa cells, hyaluronic acid, insulin, in vitro.
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