Ejaculate and type of freezing extender affect rates of fertilization of horse oocytes in vitro.
Abstract: In vitro fertilization (IVF) was performed on in vitro-matured equine oocytes in three experiments. Frozen-thawed sperm were prepared using swim-up separation and heparin treatment. In Experiment 1, fertilization was achieved with sperm from only one frozen ejaculate of four obtained from the same stallion. Within this ejaculate, fertilization rates were higher with fresh media, as compared to media held for 6-8 days before use (39.6% versus 7.3%, respectively; P<0.001). The type of bovine serum albumin used affected fertilization rates (4% versus 39.6%; P<0.001). To determine if IVF rates were influenced by factors associated with the freezing process (Experiment 2), a single ejaculate from a second stallion was frozen using eight variations in timing of steps in the freezing protocol. There were no differences among treatments in fertilization rates (range, 0-3%). In Experiment 3, fertilization rates of semen frozen in an extender containing 21.5% egg yolk were lower than fertilization rates of semen from the same ejaculate but frozen with a 3% egg-yolk extender (0% versus 15%, respectively; P<0.01). We inferred that rates of equine IVF with frozen-thawed sperm were influenced by ejaculate, the composition and age of the media used, and freezing extender. To our knowledge, this is the first report of ejaculate or extender differences affecting in vitro fertilization in this species. These factors may help to explain the great variability in fertilization rates reported with equine IVF, both among and within laboratories.
Publication Date: 2007-07-05 PubMed ID: 17614128DOI: 10.1016/j.theriogenology.2007.04.057Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research paper presents the findings of three experiments which studied how different aspects, such as the type of ejaculate, freezing extender, and media age and composition, can influence the rates of in vitro fertilization (IVF) in horses.
Experiment 1: Influence of Ejaculate and Media Age on Equine IVF Rates
- The first experiment involved in vitro fertilization on matured equine oocytes using four different frozen ejaculates from the same stallion. These were prepared through a swim-up separation technique and treated with heparin.
- Fertilization was successful with sperm from only one out of the four frozen ejaculates.
- The results significantly differed when fresh media was used as compared to media held for 6-8 days before use. The former exhibited higher fertilization rates at 39.6% while the latter displayed rates of only 7.3%.
- Furthermore, the type of bovine serum albumin used also affected the rates of fertilization. The difference between the two types was substantial with one type producing a rate as low as 4% and the other resulting in a rate as high as 39.6%.
Experiment 2: Impact of the Freezing Process on Equine IVF Rates
- The second experiment assessed if factors related to the freezing process influenced the in vitro fertilization rates.
- A single ejaculate from a second stallion was frozen using eight variations in the timing of steps in the freezing protocol.
- The results revealed no differences among the treatments in terms of fertilization rates which ranged between 0 to 3%.
Experiment 3: Effect of Freezing Extender on Equine IVF Rates
- The third experiment investigated the effect of the composition of the freezing extender on fertilization rates.
- The study found that semen frozen in an extender containing 21.5% egg yolk showed lower fertilization rates than semen from the same ejaculate but frozen with a 3% egg-yolk extender. The fertilization rates were 0% and 15% respectively.
The study concluded that factors such as the type of ejaculate, age and composition of media, and the type of freezing extender used have a significant influence on the success rates of in vitro fertilization in horses. These findings may help in explaining the variability seen in equine IVF fertilization rates both within and among different laboratories.
Cite This Article
APA
Roasa LM, Choi YH, Love CC, Romo S, Varner DD, Hinrichs K.
(2007).
Ejaculate and type of freezing extender affect rates of fertilization of horse oocytes in vitro.
Theriogenology, 68(4), 560-566.
https://doi.org/10.1016/j.theriogenology.2007.04.057 Publication
Researcher Affiliations
- Department of Physiology and Pharmacology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX 77843, USA.
MeSH Terms
- Animals
- Cell Culture Techniques
- Cryopreservation / methods
- Cryopreservation / veterinary
- Egg Yolk
- Female
- Fertilization in Vitro / methods
- Fertilization in Vitro / veterinary
- Horses / physiology
- Male
- Oocytes / physiology
- Pregnancy
- Semen Preservation / methods
- Semen Preservation / veterinary
- Serum Albumin / pharmacology
- Sperm-Ovum Interactions / physiology
Citations
This article has been cited 3 times.- de Oliveira RA, Alonso MA, Fonte JS, Fernandes CB. Equine ICSI: an update on semen perspective. Anim Reprod 2024;21(4):e20240015.
- Felix MR, Turner RM, Dobbie T, Hinrichs K. Successful in vitro fertilization in the horse: production of blastocysts and birth of foals after prolonged sperm incubation for capacitation†. Biol Reprod 2022 Dec 10;107(6):1551-1564.
- Leemans B, Stout TAE, Soom AV, Gadella BM. pH-dependent effects of procaine on equine gamete activation†. Biol Reprod 2019 Nov 21;101(5):1056-1074.
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