Authors:
T. I. Kuzmina
All-Russian Research Institute of Genetics and Breeding of Farm Animals – the branch of Federal Scientific Center for Animal Husbandry – All-Russian Research Institute of Livestock 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. Identification of the factors determining of donor’s oocyte competence to parthenogenetic development will allow developing an effective method for obtaining parthenotes to solve fundamental problems of regulating gene activity in ontogenesis, creating homozygous embryonic stem cell lines, improving the stages of cloning technology, and modeling of in vitro oocyte maturation media. The purpose of study is to evaluate the potencies of Bos taurus oocytes matured in different culture systems to cold shock-induced parthenogenesis. Methods. For oocyte maturation, culture systems of the following composition were used: 1 – TC-199 with 10 % fetal bovine serum (FBS), 50 μg/ml estradiol, 10 μg/ml luteinizing hormone, 10 μg/ml follicle-stimulating hormone; 2 – TC-199 with 10 % estrous serum of cows; 3 – TC-199 with 50 % fluid from follicles (Ø 3–8 mm); 4 – TC-199 with 50 % protein of follicular fluid (molecular weight of 65 kDa); 5 – TC-199 with 10 % FBS, 1×106 granulosa cells/ml medium; 6 – TC-199 with 10 % FBS and walls of follicles (Ø 6–8 mm); 7 – TC-199 with 10 % FBS, 1×106 granulosa cells/ml medium and walls of follicles (Ø 6–8 mm). After 24 hours of cultivation, the oocytes were activated by cold shock (exposure time 20 minutes, temperature 0...–4 °C. Results. The proportion of embryos at the stages of late morula and blastocysts from oocytes matured in system 7 was 45 % (58/129), which was significantly higher than in other systems: 1 – 28 % (39/141), P < 0.05; 2 – 31 % (42/137), P < 0.05; 3 – 25 % (33/133), P < 0.01; 4 – 18 % (25/139), P < 0.001; 5 – 31 % (41/132), P < 0.05; 6 – 33 % (43/129). The introduction of estradiol or structural components of antral follicles into the oocytes maturation medium contributed to an increase in the proportion of parthenotes at the preimplantation stages of development, including blastocysts, and a decrease in the level of degenerated embryos. Scientific novelty. A comparative morphological analysis of the potentials for parthenogenesis of bovine oocytes matured in various culture systems and activated by cold shock was carried out for the first time. Optimal systems for in vitro maturation of female gametes were proposed. Based on the analysis of the results, we recommend before induction to parthenogenesis bovine oocytes culture in media supplemented with 50 ng/ml estradiol or structural components of antral follicles producing estradiol.
Keywords: oocytes, in vitro maturation, estradiol, parthenotes, culture systems, Bos taurus.
For citation: Kuzmina T. I. Partenogeneticheskoe razvitie embrionov Bos taurus iz ootsitov, sozrevshikh v razlichnykh sistemakh kul’tivirovaniya [Partenogenetic development of Bos taurus embryos from oocytes matured in different culture systems] // Agrarian Bulletin of the Urals. 2020. No. 06 (197). Pp. 66‒72. DOI: 10.32417/1997-4868-2020-197-6-66-72. (In Russian.)
