The successful use of assisted reproduction techniques (ART) depends in part on the sperm physiological status. Several sperm selection procedures have been applied to improve quality of sperm population when using the ART. There has previously been development of a Sperm Selection Assay (SSA) for humans which is based on the attraction of capacitated sperm by chemotaxis towards progesterone (P), resulting in an enriched sperm population with an optimal physiological status similar to capacitated spermatozoa, with these cells having very little DNA fragmentation and optimal concentrations of reactive oxygen species (ROS). In the present study, the aim was to adapt the SSA for frozen-thawed stallion semen samples and evaluate the functional status of those sperm selected using the SSA procedure, and to determine whether this enriched sperm population has a greater capacity to bind to the zona pellucida of cattle oocytes. There were experimental conditions developed to conduct the SSA with stallion sperm. Using these conditions, the indexes of induced acrosome reaction, protein tyrosine phosphorylation, mitochondrial membrane potential, mitochondrial and cytoplasmic reactive oxygen species, and number of sperm bound to the zona pellucida of cattle were greater when the sperm population was selected using the SSA. Consistently, the DNA fragmentation and phospholipase C zeta indexes were less for the selected sperm. In conclusion, stallion sperm selected using chemotaxis utilizing the SSA provides a sperm population of greater quality, which when used may improve the outcomes with use of the ART.

辅助生殖技术 (ART) 的成功使用部分取决于精子的生理状态。在使用 ART 时，已经应用了几种精子选择程序来提高精子数量的质量。之前已经开发了一种用于人类的精子选择分析 (SSA)，它基于通过趋化性对孕酮 (P) 的获能精子的吸引，从而产生具有类似于获能精子的最佳生理状态的富集精子群，具有这些具有极少 DNA 断裂和最佳活性氧 (ROS) 浓度的细胞。在本研究中，目的是使 SSA 适应冻融种马精液样本，并评估使用 SSA 程序选择的精子的功能状态，并确定这种富集的精子群体是否具有更大的能力与牛卵母细胞的透明带结合。开发了一些实验条件来用种马精子进行 SSA。在这些条件下，当使用SSA选择精子种群时，诱导顶体反应、蛋白质酪氨酸磷酸化、线粒体膜电位、线粒体和细胞质活性氧以及与牛透明带结合的精子数量等指标更大。一致地，所选精子的 DNA 断裂和磷脂酶 C zeta 指数较低。总之，利用 SSA 使用趋化性选择的种马精子提供了更高质量的精子种群，使用时可以改善使用 ART 的结果。