Authors: T. I. Stanislavovich, T. I. Kuzmina
All-Russian Scientific Research Institute of Genetics and Breeding of Farm Animals – Branch of the Federal Scientific Center for Animal Husbandry – All-Russian Scientific Research Institute of Animal Husbandry named after Academician L. K. Ernst, Pushkin, Russia E-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.
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 (15 × 20 mm) 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.
Keywords: vitrification, oocyte-cumulus complex, ovarian fragments, cumulus cells, silicon dimethylglycerolate (SDMGC), Sus Scrofa Domesticus.
For citation: Stanislavovich T. I., Kuzmina T. I. Modifikatsiya etapov tekhnologii intraovarial’noy vitrifikatsii ootsitov Sus Scrofa Domesticus [Modification of the stages of the technology of intraovarian vitrification of oocytes Sus Scrofa Domesticus] // Agrarian Bulletin of the Urals. 2020. No. 08 (199). Pp. 51‒57. DOI: 10.32417/1997-4868-2020-199-8-51-57. (In Russian.)
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