ORIGINAL RESEARCH article
Front. Vet. Sci.
Sec. Animal Reproduction - Theriogenology
This article is part of the Research TopicGermplasm Conservation and Semen Banking in Livestock: Strategies, Challenges, and ApplicationsView all articles
Comparative study on in vitro fertilization, cleavage and blastocyst development of post-warmed vitrified cattle oocytes in different media
Provisionally accepted
- 1University of KwaZulu-Natal, Durban, South Africa
- 2Agricultural Research Council, Pretoria, South Africa
- 3Agricultural Research Council of South Africa (ARC-SA), Pretoria, South Africa
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This study evaluated the fertilization success and developmental competence of post-warmed vitrified cattle oocytes cultured in three distinct in vitro protocols. A total of 270 immature oocytes were randomly divided into three experimental groups (n = 90 per group), each exposed to different vitrification, warming and fertilization protocols. In Group 1; oocytes were vitrified and warmed using commercial (ART-Lab Solutions, Australia) medium, Group 2; oocytes were vitrified and warmed using TCM 199-based solution (prepared medium) and Group 3; oocytes were vitrified and warmed using Bioscience™ medium (IVF Bioscience, BO-VitriCool™ and BO-Vitri Warm™, United Kingdom). Post-warming, oocytes were matured in vitro per group: Group 1 - Vitromat-Protect™, Group 2 - TCM199, and Group 3 - BO-IVM™. Matured oocytes were then subjected to in vitro fertilization (IVF) using VitroFert™ (Group 1), prepared BO-IVF (Group 2) and BO-IVF™ (Group 3). Following 18 hours post-IVF, presumptive zygotes were washed and cultured in VitroCleave PLUSTM (Group 1), BO IVC (Group 2) and BO IVCTM (Group 3) media. Embryos were further cultured in BO IVC, BO IVCTM, or VitroBlast (Group 1) protocol with medium changes as per respective protocols. These observations were repeated 6 times. Among vitrified oocytes, BO IVF™ achieved the highest total (35.33±3.61) and normal fertilization rates (42.66±9.43), significantly higher than VitroFert™ and BO IVF. Non-vitrified oocytes showed no significant fertilization differences across media but consistently had higher cleavage rates (38.83–55.28%) than vitrified oocytes (26.50– Frontiers in veterinary sciences 30.00%). Embryos from non-vitrified oocytes cultured in BO IVC™ reached the highest morula (31.14 ± 6.17) and blastocyst (29.57 ± 6.97) development, whereas embryos from vitrified oocytes cultured in VitroCleave Plus™ yielded the highest morula and blastocyst rates (18.33 ± 15.7) compared with BO IVC™ (12.16 ± 14.14) and BO IVC (9.66 ± 15.18; p < 0.05). In conclusion, BO IVF™, and VitroCleave Plus™ medium demonstrated potential for enhancing post-warmed oocyte competence and fertilization, cleavage and blastocyst outcomes compared to other treatment groups.
Keywords: Cryopreservation, immature oocytes, Pronucleus, Cleavage, Cattle
Received: 11 Oct 2025; Accepted: 03 Nov 2025.
Copyright: © 2025 Sebopela, Mkhize, Thema and Mphaphathi. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Masindi Lottus Mphaphathi, masindim@arc.agric.za
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