Martínez-Rodríguez Carmen, Alvarez Mercedes, López-Urueña Elena, Gomes-Alves Susana, Anel-López Luis, Chamorro Cesar A, Anel Luis, de Paz Paulino
ITRA-ULE, INDEGSAL, University of León, 24071, León, Spain.
Reprod Fertil Dev. 2015 Jun;27(5):852-64. doi: 10.1071/RD13377.
Sperm motility in vitro is one of the most common predictors of fertility in male screening. We propose that a mucus-penetration assay can isolate a cellular subpopulation critical to reproductive success. To this end, a device was designed with three modules (sample, test and collection) and its conditions of use evaluated (length of mucus, incubation time, mucus medium, sperm concentration and position in relation to the horizontal). The number of spermatozoa migrating and the viability and acrosomal status of the spermatozoa not migrating were calculated. The second objective was to evaluate the qualitative parameters of the spermatozoa migrating in 1.6% polyacrylamide for 30min. The number of spermatozoa migrating and the sperm motility, viability and the acrosomal and mitochondrial status of three sperm populations (fresh, not migrating and migrating) were determined. A higher number of migrating spermatozoa were observed after 60min of incubation, but this situation adversely affected sperm quality. The methylcellulose-based test showed a significantly lower number of migrating spermatozoa than the polyacrylamide test. The position at an angle of 45° resulted in a higher number of migrating spermatozoa in the polyacrylamide-based test. The sperm counts for three consecutive assays indicated an acceptable repeatability of the method. The viability and acrosomal status of the migrating spermatozoa showed no significant changes with regard to the control when the device was placed at 45°, whereas these parameters showed lower values at 0°. The percentage of high mitochondrial membrane potential spermatozoa was significantly reduced in the population of migrating spermatozoa.
体外精子活力是男性筛查中最常见的生育能力预测指标之一。我们提出，黏液穿透试验可以分离出对生殖成功至关重要的细胞亚群。为此，设计了一种具有三个模块（样本、测试和收集）的装置，并评估了其使用条件（黏液长度、孵育时间、黏液介质、精子浓度以及相对于水平方向的位置）。计算了迁移的精子数量以及未迁移精子的活力和顶体状态。第二个目标是评估在1.6%聚丙烯酰胺中孵育30分钟的迁移精子的定性参数。测定了三个精子群体（新鲜、未迁移和迁移）的迁移精子数量、精子活力、活力以及顶体和线粒体状态。孵育60分钟后观察到迁移精子数量增加，但这种情况对精子质量有不利影响。基于甲基纤维素的试验显示迁移精子数量明显低于基于聚丙烯酰胺的试验。在基于聚丙烯酰胺的试验中，45°角的位置导致迁移精子数量增加。连续三次试验的精子计数表明该方法具有可接受的重复性。当装置置于45°时，迁移精子的活力和顶体状态与对照相比无显著变化，而在0°时这些参数值较低。迁移精子群体中高线粒体膜电位精子的百分比显著降低。
