Production of live foals via intracytoplasmic injection of lyophilized sperm and sperm extract in the horse.
Abstract: Work with lyophilized sperm helps delineate the factors required for successful fertilization. We investigated the use of lyophilized sperm in equine embryo production. In Experiment 1, sperm DNA fragmentation index was not affected by three freeze/thaw or lyophilization cycles. In Experiment 2, oocytes injected with lyophilized sperm or with sperm from a treatment in which lyophilized sperm were suspended in sperm cytoplasmic extract (SE) yielded blastocyst development rates of 0 and 28% respectively (P < 0.05). In Experiment 3, blastocyst development rate was 6-11% after injection of sperm lyophilized from fresh or frozen-thawed semen, suspended in SE. In Experiment 4, sperm lyophilized 3.5 months or 1 week previously, suspended in SE, yielded similar blastocyst rates (6 and 3% respectively). Rates of normal pregnancy after transfer were 7/10 and 5/7 for embryos from control and lyophilized sperm treatments respectively. Three pregnancies from the lyophilized sperm treatments were not terminated, resulting in two healthy foals. Parentage testing determined that one foal originated from the lyophilized sperm; the other was the offspring of the stallion providing the sperm extract. Further testing indicated that two of five additional embryos in the lyophilized sperm treatment originated from the stallion providing the sperm extract. We conclude that both lyophilized stallion sperm and stallion sperm processed by multiple unprotected freeze-thaw cycles (as for sperm extract) can support production of viable foals. To the best of our knowledge, this is the first report on production of live offspring by fertilization with lyophilized sperm in a non-laboratory animal species.
Publication Date: 2011-08-16 PubMed ID: 21846810DOI: 10.1530/REP-11-0145Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research explores how freeze-drying sperm can be used in horse breeding to generate live offspring. The study found that both lyophilized (freeze-dried) and multiple freeze-thaw processed stallion sperm can support the creation of viable foals.
Methodology
- The study consisted of four experiments. These experiments were designed to understand the effects of the freeze/thaw cycle on sperm DNA fragmentation, efficiency of lyophilized sperm in embryo production, and the blastocyst development rate of embryos fertilized with fresh or frozen-thawed lyophilized sperm suspended in sperm cytoplasmic extract (SE).
- Lyophilization, also known as freeze-drying, is the process used in preserving sperm. The term covers multiple freeze-thaw cycles and storing sperm for long periods while retaining as much fertility as possible.
Key Findings
- The first experiment revealed that the sperm DNA fragmentation index was not affected by three freeze/thaw or lyophilization cycles. This is significant because DNA fragmentation can lead to impaired fertility, so demonstrating that freeze-drying doesn’t harm the DNA offers hope for its use in fertilization.
- In the second experiment, oocytes, or egg cells, were injected with either lyophilized sperm or sperm from a treatment in which lyophilized sperm were suspended in SE. The resulting blastocyst development rates were 0 and 28% respectively, suggesting better results when adding SE.
- The third experiment showed that using lyophilized sperm from fresh or frozen-thawed semen, when suspended in SE, yielded a blastocyst development rate of 6-11%.
- In the fourth experiment, similar blastocyst rates (6 and 3%) were found for sperm that was lyophilized either 3.5 months or one week prior and then suspended in SE. This indicates the long-term storage potential of lyophilized sperm.
- The study also documented normal pregnancy rates after transfer of embryos. The embryos either originated from normal processes (control) or from lyophilized sperm treatments. Seven out of ten control embryos and five out of seven from the lyophilized sperm treatments resulted in a normal pregnancy.
- Among the successful pregnancies from the lyophilized sperm treatments, three were not terminated and resulted in two healthy foals. DNA tests confirmed one foal originated from the lyophilized sperm, while the second was the offspring of the stallion whose sperm extract was used.
Conclusion
- The study concluded that live offspring can be successfully produced through fertilization with lyophilized sperm and freeze-thaw processed sperm in horses. This signifies a significant breakthrough in animal reproduction science.
- This research is the first to document the production of live offspring by fertilization using lyophilized sperm in any non-laboratory animal species.
Cite This Article
APA
Choi YH, Varner DD, Love CC, Hartman DL, Hinrichs K.
(2011).
Production of live foals via intracytoplasmic injection of lyophilized sperm and sperm extract in the horse.
Reproduction, 142(4), 529-538.
https://doi.org/10.1530/REP-11-0145 Publication
Researcher Affiliations
- Department of Veterinary Physiology and Pharmacology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, Texas 77843-4466, USA.
MeSH Terms
- Animals
- Blastocyst / cytology
- Cell Survival / physiology
- DNA Fragmentation
- Embryo, Mammalian / physiology
- Embryonic Development / physiology
- Female
- Freeze Drying
- Horses / embryology
- Horses / physiology
- Live Birth / veterinary
- Male
- Pregnancy
- Pregnancy Outcome
- Pregnancy, Animal / physiology
- Sperm Injections, Intracytoplasmic / methods
Citations
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