Abstract
Abstract. Intraovarian vitrification of oocytes of farm animals today is a very promising direction. The composition of cryoprotective solutions requires further improvement. Vitrification of ovarian fragments (FО) allows tо save primordial follicles, a structure that is more stable, due to the lack of follicular fluid, to destructive processes provoked by ultra-low temperatures.The high reproductive potential of pigs is due to their physiological characteristics of growth and development. Silicon is a minor trace element in its content; it is increasingly used in pharmacology, veterinary medicine, and clinical practice. Nitrogen silicon dimethylglycerolate (SDMGC) are antimicrobial, wound healing and anti-inflammatory in nature. The presence of a compact cumulus surrounding the oocyte is one of the important indicators of the reliability of the female gamete for extrafollicular maturation. Purpose of this study: to was to evaluate the effect of introducing 0.2 % silicon dimethylglycerolate into cryoprotective solutions during intraovarian vitrification and the medium for maturation of devitrified oocyte-cumulus complexes of pigs on the morphology of cumulus cells and the state of nuclear material. Ovarian fragments of pigs (15mmx 20mm) to vitrification are exposed prior for 25 minutes and 15 minutes direct in cryoprotective solutions of the following composition: CPA 1 – 7.5 % ethylene glycol (EG), 7.5 % dimethyl sulfoxide (DMSO), 65 % phosphate-buffered saline (PBS), 20 % fetal bovine serum (PBS); CPA 2 – 20 % EG, 20 % DMSO, 60 % FSB, 0.5 mol / L sucrose. The experimental group was treated for 10 minutes in a solution of phosphate-saline buffer with 0.2 % SDMGC. Hoechst 33258 was used to evaluate chromatin status, samples were analyzed using a ZEISS Axio Imager Imager A 2 m microscope.The result. Incubation of ovarian fragments of pigs in a solution containing 0.2 % SDMGC before vitrification, as well as the introduction of 0.2 % SDMGC in the composition of oocyte maturation media, had a positive effect on the morphology of cumulus cells (degree of expansion) after the thawing procedure. The proportion of compact cumulus devitrified cells pretreated with 0.2 % SDMGC increased compared with the control group (64 % vs 56 %, P < 0.05 (χ2-test). The level of oocytes with cumulus is in a high degree of expansion after 44 hours of cultivation with 0.2 % SDMGC in the experimental group was 75 % vs 54 % in the control group, P < 0.05 (χ2-test). The use of SDMGC in the cultivation of devitrified oocyte-cumulus complexes of pigs for 44 hours increased the proportion of matured oocytes from 41 % in the control group to 64 % in the experimental group P < 0.05 (χ2-test). The scientific novelty: the stages of the technology of intraovarian vitrification of porcine oocytes by means of preventive incubation of ovarian fragments in a solution of 0.2 % SDMGC and its introduction into the medium for culturing oocyte-cumulus complexes were modernized.
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