Singina GN, Chinarov RYu, Shedova EN.
Animal Husbandry and Fodder Production. 2025. Vol. 108. No. 4. Р. 31-40.
doi:10.33284/2658-3135-108-4-31
Original article
Optimization of in vitro development conditions for OPU/IVP bovine embryos: the effect
of medium replacement
Galina N Singina1, Roman Yu Chinarov2, Ekaterina N Shedova3
1,2,3Federal Research Center for Animal Husbandry named after Academy Member LK Ernst, Dubrovitsy, Russia
1g_singina@mail.ru, https://orcid.org/0000-0003-0198-9757
2roman_chinarov@mail.ru, https://orcid.org/0000-0001-6511-5341
3shedvek@yandex.ru, https://orcid.org/0000-0002-9642-2384
Abstract. Oocyte retrieval from follicles under ultrasound guidance (OPU) and the subsequent in vitro embryo production (IVP) represent a modern biotechnological approach in cattle breeding that enables the generation of a greater number of offspring from genetically valuable animals. A stage of the OPU/IVP procedure, the optimization of which can substantially improve its efficiency, is the culture of matured oocytes after in vitro fertilization up to the blastocyst (Bl) stage. The aim of this study was to investigate the effect of culture medium renewal conditions on the development and quality of IVP embryos during in vitro culture (IVC). OPU-derived bovine oocytes, following in vitro maturation and fertilization, were transferred into BO-IVC medium for culture and embryonic development. The effects of not refreshing the IVC medium during the 7-day embryonic development period (NRM) and completely refreshed medium (CRM) on day 3 of IVC were compared. The results demonstrated that, compared with NRM, CRM increased blastocyst yield from 19.3±0.7% to 31.7±2.0% (P=0.005). In addition, a higher proportion of excellent- and good-quality blastocysts was observed in the CRM group compared with the NRM group (53.6±5.9% vs. 22.9±3.4%, P=0.011). Therefore, the in vitro developmental performance of bovine embryos in BO-IVC medium appears to depend on the medium renewal regimen. To achieve higher blastocyst yield and quality, a complete medium replacement on the third day of culture is recommended.
Keywords: cattle, owum pick up, in vitro embryo production, embryo culture medium
Acknowledgments: the work was performed in accordance to the plan of research works of the FGBNU FRC VIZh named after academician LK Ernst (No FGGN-2025-0008).
For citation: Singina GN, Chinarov RYu, Shedova EN. Optimization of in vitro development conditions for OPU/IVP bovine embryos: the effect of medium replacement. Animal Husbandry and Fodder Production. 2025;108(4):31-40. (In Russ.). https://doi.org/10.33284/2658-3135-108-4-31
References
- Zinovieva NA, Pozyabin SV, Chinarov RYu. Assisted reproductive technologies: the history and role in the development of genetic technologies in cattle (review). Sel'skokhozyaistvennaya Biologiya [Agricultural Biology]. 2020;55(2):225-242. doi: 10.15389/agrobiology.2020.2.225rus doi: 10.15389/agrobiology.2020.2.225eng
- Singina GN, Bagirov VA, Danch SS, Taradainik TE, Dotsev AV, Zinovieva In vitro production of interspecies hybrid embryos of cattle (Bos taurus) and wisent (Bison bonasus). Sel'skokhozyaistvennaya Biologiya [Agricultural Biology]. 2016;51(6):824-829. doi: 10.15389/agrobiology.2016.6.824rus doi: 10.15389/agrobiology.2016.6.824eng
- Singina GN, Chinarov RYu, Shedova EN, Lukanina VA, Zhukova AS. Results of oocyte isolation and obtaining IVP embryos from heifers of Istoben breed depending on OPU regimen. Animal Husbandry and Fodder Production. 2024;107(4):283-294. doi: 10.33284/2658-3135-107-4-283
- Babenkov VYu, Chimidova NV, Khakhlinov AI, Ubushieva AV, Manjiev VI. The role of reproductive biotechnologies in the reproduction and preservation of the gene pool of rare and endangered breeds of cattle. Animal Husbandry and Fodder Production. 2023;106(1):67-76. doi: 10.33284/2658-3135-106-1-67
- Aguila L, Treulen F, Therrien J, Felmer R, Valdivia M, Smith LC. Oocyte selection for in vitro embryo production in bovine species: noninvasive approaches for new challenges of oocyte competence. 2020;10(12):2196. doi: 10.3390/ani10122196
- Amaral TF, de Grazia JGV, Martinhao LAG, De Col F, Siqueira LGB, Viana JHM, Hansen PJ. Actions of CSF2 and DKK1 on bovine embryo development and pregnancy outcomes are affected by composition of embryo culture medium. Scientific Reports. 2022;12(1):7503. doi: 10.1038/s41598-022-11447-7
- Bó GA, Mapletoft RJ. Evaluation and classification of bovine embryos. Animal Reproduction. 2013;10(3):344-348.
- Ferré LB, Kjelland ME, Strøbech LB, Hyttel P, Mermillod P, Ross PJ. Review: Recent advances in bovine in vitro embryo production: reproductive biotechnology history and methods. Animal. 2020a;14(5):991-1004. doi: 10.1017/S1751731119002775
- Ferré LB, Kjelland ME, Taiyeb AM, Campos-Chillon F, Ross PJ. Recent progress in bovine in vitro-derived embryo cryotolerance: impact of in vitro culture systems, advances in cryopreservation and future Reproduction in Domestic Animals. 2020b;55(6):659-676. doi: 10.1111/rda.13667
- Fukui Y, Lee ES, Araki N. Effect of medium renewal during culture in two different culture systems on development to blastocysts from in vitro produced early bovine embryos. Journal of Animal Science. 1996;74(11):2752-2758. doi: 10.2527/1996.74112752x
- Gutierrez-Castillo E, Ming H, Foster B, Gatenby L, Mak CK, Pinto C, Bondioli K, Jiang Z. Effect of vitrification on global gene expression dynamics of bovine elongating embryos. Reproduction Fertility Development. 2021;33(5):338-348. doi: 10.1071/RD20285
- Hansen PJ. Review: Some challenges and unrealized opportunities toward widespread use of the in vitro-produced embryo in cattle production. Animal.2023;17(S1):100745. doi: 10.1016/j.animal.2023.100745
- Ikeda K, Takahashi Y, Katagiri S. Effect of medium change on the development of in vitro matured and fertilized bovine oocytes cultured in medium containing amino acids. The Journal of Veterinary Medical Science. 2000;62(1):121-123. doi: 10.1292/jvms.62.121
- Lonergan P, Fair T, Forde N, Rizos D. Embryo development in dairy cattle. Theriogenology. 2016;86(1):270-277. doi: 10.1016/j.theriogenology.2016.04.040
- Mullaart E, Ludema G, Zijlstra A, Veldhuisen J. 64 No difference in health and fertility characteristics between offspring arising from in vitro production or multiple ovulation embryo transfer. Reproduction Fertility and Development. 2021;34(2):267-268. doi: 10.1071/RDv34n2Ab64
- Nielsen JMK, Wrenzycki C, Hyttel P, Poppicht F, Strøbech L. New culture media affects blastocyst development and gene expression levels in in vitro-produced bovine embryos. Reproduction Fertility Development. 2014;27(1):206-207. doi: 10.1071/RDv27n1Ab234
- Qu P, Qing S, Liu R, Qin H, Wang W, Qiao F, Ge H, Liu J, Zhang Y, Cui W, Wang Y. Effects of embryo-derived exosomes on the development of bovine cloned embryos. PLoS ONE. 2017;12(3):e0174535. doi: 10.1371/journal.pone.0174535
- Sanches BV, Zangirolamo AF, Seneda MM. Intensive use of IVF by large-scale dairy programs. Animal Reproduction. 2019;16(3): 394-401. doi: 10.21451/1984-3143-AR2019-0058
- Sciorio R, Smith GD. Embryo culture at a reduced oxygen concentration of 5 %: a mini review. Zygote. 2019;27(6):355-361. doi: 10.1017/S0967199419000522
- Takahashi M, Keicho K, Takahashi H, Ogawa H, Schultz RM, Okano A. Effect of oxidative stress on development and DNA damage in in-vitro cultured bovine embryos by comet assay. Theriogenology. 2000;54(1):137-145. doi: 10.1016/s0093-691x(00)00332-0
- Viana J. IETS 2017 Statistics of embryo production and transfer in domestic farm animals. Embryo Technology Newsletter. 2018;36(4):8-25.
- Viana J. IETS 2023 Statistics of embryo production and transfer in domestic farm animals. Embryo Technology Newsletter. 2024;43(4):31-46.
Information about the authors:
Galina N Singina, Cand. Sci. (Biology), Head of the Laboratory of Embryonic Technologies, Federal Research Center for Animal Husbandry named after Academy Member LK Ernst, 60 Dubrovitsy village, Podolsk City district, Moscow region, 142132, mobile: 8-9851434552.
Roman Yu Chinarov, Cand. Sci. (Veterinary), Senior Researcher, Head of the Laboratory of Embryonic Technologies, Federal Research Center for Animal Husbandry named after Academy Member LK Ernst, 60 Dubrovitsy village, Podolsk City district, Moscow region, 142132, mobile: 8-9153899065.
Ekaterina N Shedova, Researcher, Head of the Laboratory of Embryonic Technologies, Federal Research Center for Animal Husbandry named after Academy Member LK Ernst, 60 Dubrovitsy village, Podolsk City district, Moscow region, 142132, mobile: 8-9167117673.
The article was submitted 06.10.2025; approved after reviewing 28.11.2025; accepted for publication 15.12.2025.
Download