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Dog sperm cryopreservation using cryovials and different dilution steps
S Ibrahim1, N A Hamad Talha1, J Cho2
1Laboratory of Theriogenology and Reproductive Biotechnology, College of Veterinary Medicine and Bio-safety Research Institute, Jeonbuk National University, Iksan 54596, Republic of Korea. Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, Sudan University of Science and Technology, Khartoum 11111, Sudan.
A new cryovial method for dog sperm freezing is more efficient than traditional straw freezing. This technique preserves sperm quality and reduces oxidative stress, offering a simpler and effective cryopreservation solution.
Area of Science:
- Veterinary Reproduction
- Cryobiology
- Canine Genetics
Background:
- Conventional dog sperm freezing uses straws, requiring two-step dilution and lengthy equilibration.
- This process can be time-consuming and may impact sperm viability.
Purpose of the Study:
- To develop a more efficient sperm cryopreservation method for dogs.
- To evaluate the efficacy of using cryovials with a simplified dilution protocol.
Main Methods:
- Three protocols using 0.5 mL cryovials with varying cooling and dilution steps were tested on dog spermatozoa.
- Spermatozoa were cooled and frozen using different combinations of extenders and temperatures, including a deep freezer and liquid nitrogen (LN2).
- A control group used the conventional straw method with two-step dilution.
Main Results:
- Cryovial methods (Groups 2 and 3) showed high progressive motility and acrosome integrity, comparable to the conventional method.
- Group 3 exhibited superior viability, while Group 2 demonstrated significantly lower reactive oxygen species (ROS) and phosphatidylserine (PS) translocation.
- Gene expression analysis revealed higher levels of anti-apoptotic BCL2 and SMCP in Group 2, and lower BAX expression.
Conclusions:
- Cryovial freezing with one-step dilution and deep freezer storage (Group 2) is a simple and effective method for dog sperm cryopreservation.
- This optimized method offers improved sperm quality and reduced oxidative damage compared to conventional techniques.
