E. V. Kornienko1, M. A. Ikonopistseva2, G. P. Malenko1
1Center for Experimental Embryology and Reproductive Biotechnology, ul. Kostyakova, 12, str. 4, Moskva, 127422, Russian Federation
2Lomonosov Moscow State University, Leninskie gory, 1, str. 12, Moskva, 119991, Russian Federation
Abstract. The goal of the current work was to enhance the effectiveness of vitrification in triacetate cellulose hollow fiber at cryopreservation of in vitro matured bovine oocytes. Experiments were carried out in 2017–2018. For vitrification/rewarming of oocytes the basic medium TLP-HEPES with 20% of bovine fetal serum was used, prepared either with (TH20) or without (TH20–) calcium salts. In the case of TH20– medium, only ethylene glycol was used as permeating cryoprotectant. In the case of TH20 medium, we used both ethylene glycol and dimethyl sulfoxide. In vitro matured bovine oocytes demonstrated high survival ((80.3 ± 1.5)% and (79.0 ± 8.5)%) and fertilization ((83.9 ± 7.9)% and (84.4 ± 18.1)%) rates in groups TH20 and TH20–, correspondingly. In TH20– group, blastocysts were obtained only in 50% of repeats (6 repeats in total, 140 oocytes); in TH20 group they were obtained in 66.7% of repeats (3 repeats in total, 65 oocytes). On the 7th and the 10th days of culture in group TH20, the output of embryos on the blastocyst stage was (3.1 ± 4.4)% (1/48) and (7.8 ± 6.6)% (3/48), correspondingly. In group TH20– the output was (1.9 ± 3.2)% (1/51) and (5.9 ± 0.6)% (3/51), respectively. These results are comparable with the vitrification of in vitro matured bovine oocytes using various open-type devices. Blastocyst hatching was observed only in group TH20– (3/3), which indicates a possibility of the protocol modification via decreasing of Ca2+ effect during incubation in vitrification/rewarming solutions. In the current work for the first time the effective use of the method of the group vitrification in triacetate cellulose hollow fibers of in vitro matured bovine oocytes was demonstrated.
Keywords: oocyte; vitrification; cattle; triacetate cellulose hollow fibers.
Author Details: E. V. Kornienko, research fellow (е-mail: Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.); M. A. Ikonopistseva, master’s student; G. P. Malenko, D. Sc. (Biol.), head of division.
For citation: Kornienko E. V., Ikonopistseva M. A., Malenko G. P. Vitrification of In Vitro Matured Bovine Oocytes in Triacetate Cellulose Hollow Fibers. Dostizheniya nauki i tekhniki APK. 2018. Vol. 32. No. 9. Pp. 84–88 (in Russ.). DOI: 10.24411/0235-2451-2018-10920.