Mesenchymal stem cells (MSCs) are believed to be an integral part of regenerative medicine. Therefore, the repair of injured sperm could be a determining factor for improving the fertility of canine using frozen semen. In addition, the chemical additives used for supplementation have been also associated with the issues of cytotoxicity 8, 18.
The reasons behind the lower fertility of frozen semen may be the primary or secondary damaged sperm that requires immediate regeneration and recovery. However, the rates of whelping using cryopreserved semen (50.0–70.8%) are still lower in comparison with the fresh semen (81.8–83.7%) 17.
These changes ultimately contribute to an overall reduction in the fertility of sperm.Ĭurrent protective modifications employed to minimize the above mentioned damaging effects involve the use of different types of extenders 9, variations in the freezing protocols 10, and supplementation of extenders with nutrients or antioxidants 11, 12, 13, 14, 15, 16. In addition, the activation of apoptotic pathways results in the fragmentation of sperm cell DNA 8. The sperm that survives during freezing procedure suffers a reduction in fertility 6 and this has been linked with damage that adversely affects viability, motility, plasma membrane, acrosome and chromatin material 7. Sperm undergoes certain detrimental changes at the structural and molecular levels as a result of thermal, mechanical, osmotic, and oxidative damage 4, 5. During freezing the quality-related traits of sperm are highly compromised 3. In addition, it has greatly benefited the animal -based industry by reducing the cost and the stress associated with transportation, overcoming the quarantine restrictions and breeding issues (e.g., aggressive behavior and size issues) 2.ĭespite many attempts to improve freezing agents and techniques, reduction in semen quality still remains the major issue associated with freezing procedures. It can facilitate the storage of the genetic material for an extended period, conserve the elite individual’s fertility and serve as a useful tool for preserving the endangered species. Semen cryopreservation is regarded as the most important step of artificial insemination which is the most widely adopted assisted reproductive technology (ART) in canine practice 1. At an appropriate concentration, Ad -MSCs significantly improve the quality of post-thaw dog sperm. The results confirm that canine Ad -MSCs can effectively preserve the quality of frozen-thawed sperm by a reduction in cryoinjury. Protein expression of ANX1, H 3, and FN1 was also statistically more in Group 1 than in Control. Additionally, Group 1 showed significantly higher expression levels of genes related to the repair of membranes ( ANX1, dysferlin DYSF, and fibronectin FN1) and chromatin material ( H3 and HMGB). Group 1 exhibited significantly higher post-thaw motility, live sperm, intact plasma membrane and normal acrosomes than the other groups. The washed pooled ejaculates were diluted with buffer 2 (extender) supplemented without Ad -MSCs (Control), with 2.5 × 10 6 Ad -MSCs/mL (Group 1) or with 5 × 10 6 Ad -MSCs/mL (Group 2). Semen was collected from four healthy dogs by digital manipulation.
#Minitube canine extender skin#
Canine Ad -MSCs were selected on the basis of the significantly higher gene expression for different proteins actively involved in the cell repair including annexin 1 ( ANX1), histone H3 ( H3) and high mobility group B ( HMGB) protein compared to skin fibroblasts. The objective of this study was to examine the potential of canine adipose -derived mesenchymal stem cells (Ad -MSCs) for providing protection to the dog sperm against cryo-damage. Cryopreservation procedures negatively affect the quality traits of sperm, causing certain changes at structural and molecular levels due to thermal, mechanical, osmotic, and oxidative damage.
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