Single and double layer centrifugation improve the quality of cryopreserved bovine sperm from poor quality ejaculates

Alessia Gloria; Augusto Carluccio; Laura Wegher; Domenico Robbe; Giovanni Befacchia; Alberto Contri

摘要

Background:Density gradient centrifugation was reported as a technique of semen preparation in assisted reproductive techniques in humans and animals.This technique was found to be efficient in improving semen quality after harmful techniques such as cryopreservation.Recently a modified technique,single layer centrifugation,was proposed as a technique providing a large amount of high quality spermatozoa,and this treatment was performed before conservation.Single layer centrifugation has been studied prevalently in stallions and in boars,but limited data were available for bulls.Occasionally bulls are known to experience a transient reduction in semen quality,thus techniques that allow improvement in semen quality could be applied in this context.The aim of this study was the evaluation of single layer and double layer centrifugation by the use of iodixanol,compared with conventional centrifugation and non-centrifuged semen,on the sperm characteristics during the cryopreservation process in bulls with normal and poor semen quality.Results:Single layer centrifugation and double layer centrifugation both significantly increased the percentage of normal spermatozoa and decreased the percentage of non-sperm cells in poor quality samples,while both were ineffective in those of normal quality.Sperm characteristics in poor quality samples increased after single layer centrifugation and double layer centrifugation,reaching values similar to those recorded in normal samples,and this trend is maintained after equilibration and after cryopreservation.On the other hand,SLC and DLC resulted in a consistent reduction in the spermatozoa recovered,and this resulted in a reduction of the absolute amount of spermatozoa cryopreserved in the normal samples,without a clear improvement in sperm characteristics in this type of sample.Conclusions:These data suggested that both SLC and DLC could be performed in practice,but their application should be limited to the cases in which the quality of the spermatozoa recovered is more important than the total amount of spermatozoa.

机译：背景：密度梯度离心技术是人类和动物辅助生殖技术中精液制备的一种技术。经过冷冻保存等有害技术后，发现该技术可有效提高精液质量。最近，单层离心技术得到了改进。作为提供大量高质量精子的技术提出的，并且在保存之前就进行了这种处理。在种马和公猪中普遍进行了单层离心研究，但是对于公牛只有有限的数据。已知公牛会经历短暂的精液质量的降低，因此可以在这种情况下应用改善精液质量的技术。本研究的目的是评估碘克沙醇与常规离心法和非离心精液相比单层和双层离心的效果。冷冻过程中精子的特性结果：单层离心和双层离心均显着提高了劣质样品中正常精子的百分比，降低了非精子细胞的百分比，而对正常质量的样品均无效。单层离心和双层离心后，劣质样品的精子特性增加，达到与正常样品中记录的值相似的水平，平衡后和冷冻保存后这种趋势得以保持。另一方面，SLC和DLC导致一致回收的精子减少，这导致正常样品中冷冻保存的精子的绝对量减少，而这类样品的精子特性没有明显改善。结论：这些数据表明可以同时进行SLC和DLC在实践中，但其应用应仅限于具有质量要求的情况y回收的精子比精子的总量更重要。

Single and double layer centrifugation improve the quality of cryopreserved bovine sperm from poor quality ejaculates

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