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Comparison of semen preservation methods and in vitro fertilisation embryo quality in Simmental cattle |
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Kai Hu1, Fei Huang2, Hong Chen2, Hui-Min Qu1, Xiao-Peng Li1, Xue-Yan Wang1, Jie-Ru Wang2, Peng Niu2, Di Fang1, Qinghua Gao1,2,* |
1College of Animal Science and Technology, Aral, China
2Tarim University, Aral, China
3College of Life Science and Technology, Aral, China
4Key Laboratory of Tarim Animal Husbandry Science and Technology, Aral, China |
Correspondence:
Qinghua Gao, Tel: +86-13289976139, Email: gqhdky@126.com
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Received: 5 June 2025 • Revised: 26 July 2025 • Accepted: 25 November 2025 |
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| Abstract |
Objective
Semen quality plays a crucial role in embryo production in cattle. This study aims to compare the effects of 0°C refrigerated semen and liquid nitrogen-frozen semen on the quality of in vitro fertilised embryos in Simmental cattle.
Methods
Semen from six bulls was split equally into two groups. One was diluted with cold storage solution and stored at 0°C for 72 h; the other with cryopreservation solution and preserved in liquid nitrogen for 72 h. Post-storage, sperm quality (motility, progressive motility via CASA, acrosome/plasma membrane integrity) was assessed, along with MDA, ROS, ATP, and MMP levels. For IVF, oocytes from Simmental cows (>50 per group) were inseminated with the two semen groups. At 45 h post-IVF, embryo cleavage rate was recorded; on day 8, blastocyst rate was determined. Embryos then underwent BrdU staining (proliferation) and TUNEL staining (apoptosis), with proliferation and apoptosis levels quantified via marker genes.
Results
The results showed that compared with liquid nitrogen-frozen semen, 0°C refrigerated semen exhibited significantly higher sperm motility, forward motility rate, acrosome integrity, and plasma membrane integrity (p < 0.05). In vitro fertilised embryos also exhibited significant increases in quality and quantity (cleavage rate and blastocyst rate) (p < 0.05), and embryos derived from 0°C refrigerated semen contained significantly more proliferating cells (p < 0.05) and fewer apoptotic cells (p < 0.05) than those from frozen semen.
Conclusion
These findings suggest that 0°C refrigerated semen is superior to liquid nitrogen frozen semen in terms of sperm quality parameters and in vitro embryo production.
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| Keywords:
Semen; 0℃ refrigerated; Liquid nitrogen-frozen; Sperm quality; In vitro fertilisation; Embryo quality |
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