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Singina G.N., Shedova E.N., Lopukhov A.V., Zhukova A.S. IGF1 and FGF2 particular effects on the quality of maturing cattle (Bos taurus taurus) oocytes and their competence to embryonic development in vitro. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2025, Vol. 60, № 6, p. 1085-1096.
(how to cite this article)

doi: 10.15389/agrobiology.2025.6.1085eng

UDC: 636.2: 91.39

Acknowledgements:
Supported financially by the Russian Science Foundation, project No. 24-16-00261.

 

IGF1 AND FGF2 PARTICULAR EFFECTS ON THE QUALITY OF MATURING CATTLE (Bos taurus taurus) OOCYTES AND THEIR COMPETENCE TO EMBRYONIC DEVELOPMENT in vitro

G.N. Singina✉, E.N. Shedova, A.V. Lopukhov, A.S. Zhukova

Ernst Federal Research Center for Animal Husbandry, 60, pos. Dubrovitsy, Podolsk District, Moscow Province, 142132 Russia, e-mail g_singina@mail.ru (✉ corresponding author),shedvek@yandex.ru, vubi_myaso@mail.ru,
anastasia.s.belyaeva@gmail.com

ORCID:
Singina G.N. orcid.org/0000-0003-0198-9757
Lopukhov A.V. orcid.org/0000-0002-1284-1486
Shedova E.N. orcid.org/0000-0002-9642-2384
Zhukova A.S. orcid.org/0000-0003-1155-014X

Final revision received September 30, 2025
Accepted November 26, 2025

In vitro maturation (IVM) is an essential component of in vitro embryo production (IVP). The conditions under which IVM is performed critically determine the quality of oocytes and the resulting IVP embryos; however, current maturation protocols remain suboptimal and require further improvement. In the present study, we demonstrate for the first time that bovine oocytes matured in vitro in the presence of IGF1 or FGF2, and subsequently subjected to prolonged cytokine-free culture, differ in their ability to develop to the blastocyst stage following in vitro fertilization (IVF). The aim of this work was to comparatively assess the effects of two cytokines, the insulin-like growth factor 1 (IGF1) and fibroblast growth factor 2 (FGF2) when added to the oocyte in vitro maturation medium, focusing on oocyte quality, embryonic developmental potential, and resistance to age-related alterations. Ovaries from sexually mature cows (Bos taurus taurus) were collected post-mortem and transported to the laboratory in physiological saline within 4-6 h at a temperature not below 28 °C. Cumulus–oocyte complexes (COCs) were mechanically isolated from visible follicles by dissecting follicular walls with a scalpel blade, followed by retrieval and washing of the collected complexes. Groups of 20-25 selected COCs were matured in vitro for 20 h in control TC-199C medium supplemented with HEPES (25 mM), sodium pyruvate (0.5 mM), follicle-stimulating and luteinizing hormones (10 mg/ml each), epidermal growth factor (20 ng/ml), fetal bovine serum (10 %), and gentamicin (50 mg/ml). Experimental groups received identical medium supplemented with IGF1 (20 ng/ml) or FGF2 (40 ng/ml). After IVM, a subset of matured COCs was transferred into an aging medium (TC-199 with 10 % serum) and cultured for an additional 12 or 24 h. Immediately after IVM and after 24 h of aging, apoptosis was evaluated using the TUNEL assay; in the former case, meiotic staging was additionally performed. Nuclear maturation was quantified as the rate of oocytes at specific meiotic stages, while apoptosis was assessed as the rate of TUNEL-positive metaphase II (MII) oocytes among the total MII population. A portion of matured oocytes and oocytes aged for 12 h underwent IVF and subsequent embryo culture. Blastocyst quality was assessed using cytological preparations. Oocyte maturation rate (progression to meiosis II metaphase, MII) did not differ between groups, ranging from 76.5 % to 85.2 %. However, IGF1 and FGF2 treatments reduced apoptotic degeneration in MII oocytes: from 22.5±1.4 % in control to 7.6±1.4 % and 12.3±0.2 % after 20 h IVM, respectively (p < 0.001), and from 40.8±1.8 % in control to 23.2±1.0 % and 19.8±0.78 % after 24 h aging respectively (p < 0.001). For the 20-h IVM period, IGF1 demonstrated a significantly stronger protective effect than FGF2 (p < 0.05). In addition, IGF1 enhanced in vitro embryonic development: blastocyst rate following in vitro fertilization of matured (20 h IVM) and aged (12 h) oocytes reached 37.0±5.50 % and 23.3±2.9 %, exceeding control values by 16 % and 12 %, respectively (p < 0.05). Furthermore, IGF1 improved blastocyst quality by reducing the apoptosis rate in blastocysts (p < 0.05). FGF2 exerted a positive effect only on oocytes fertilized immediately after IVM, with no benefit observed in aged oocytes. Thus, IGF1 exposure during IVM enhances oocyte quality, including the resistance of bovine oocytes to subsequent age-related, and may serve as an effective means of improving IVP efficiency in cattle.

Keywords: insulin-like growth factor 1, fibroblast growth factor 2, in vitro maturation, aging, embryonic development, apoptosis, cattle.

 

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