Gimeno Brenda Florencia, Bariani María Victoria, Laiz-Quiroga Lucía, Martínez-León Eduardo, Von-Meyeren Micaela, Rey Osvaldo, Mutto Adrián Ángel, Osycka-Salut Claudia Elena
Laboratorio de Biotecnologías Reproductivas y Mejoramiento Genético Animal, Instituto de Investigaciones Biotecnológicas, Universidad Nacional de San Martín (UNSAM), Campus Miguelete, Avenida 25 de Mayo y Francia, San Martín, Buenos Aires, CP 1650, Argentina.
Signaling and Cancer Laboratory, Consejo Nacional de Investigaciones Científicas y Técnicas, Instituto de Inmunología, Genética y Metabolismo, Facultad de Farmacia y Bioquímica, Hospital de Clínicas "José de San Martín", Universidad de Buenos Aires, Ciudad Autónoma de Buenos Aires (CABA), CP 1120, Argentina.
Animals (Basel). 2021 Jan 3;11(1):74. doi: 10.3390/ani11010074.
Cryopreservation by negatively affecting sperm quality decreases the efficiency of assisted reproduction techniques (ARTs). Thus, we first evaluated sperm motility at different conditions for the manipulation of equine cryopreserved spermatozoa. Higher motility was observed when spermatozoa were incubated for 30 min at 30 × 10/mL compared to lower concentrations ( < 0.05) and when a short centrifugation at 200× was performed ( < 0.05). Moreover, because sperm suitable for oocyte fertilization is released from oviduct epithelial cells (OECs), in response to the capacitation process, we established an in vitro OEC culture model to select a sperm population with potential fertilizing capacity in this species. We demonstrated E-cadherin and cytokeratin expression in cultures of OECs obtained. When sperm-OEC cocultures were performed, the attached spermatozoa were motile and presented an intact acrosome, suggesting a selection by the oviductal model. When co-cultures were incubated in capacitating conditions a greater number of alive ( < 0.05), capacitated ( < 0.05), with progressive motility ( < 0.05) and with the intact acrosome sperm population was observed ( < 0.05) suggesting that the sperm population released from OECs in vitro presents potential fertilizing capacity. Improvements in handling and selection of cryopreserved sperm would improve efficiencies in ARTs allowing the use of a population of higher-quality sperm.
冷冻保存会对精子质量产生负面影响,从而降低辅助生殖技术(ARTs)的效率。因此,我们首先评估了在不同条件下处理马冷冻精子时的精子活力。与较低浓度(<0.05)相比,当精子在30×10/mL的浓度下孵育30分钟时,观察到更高的活力;并且当进行200×的短时间离心时(<0.05),也观察到更高的活力。此外,由于适合卵母细胞受精的精子是从输卵管上皮细胞(OECs)释放出来的,以响应获能过程,我们建立了一种体外OEC培养模型,以选择该物种中具有潜在受精能力的精子群体。我们在获得的OEC培养物中证实了E-钙黏蛋白和细胞角蛋白的表达。当进行精子-OEC共培养时,附着的精子具有活力且顶体完整,这表明输卵管模型进行了筛选。当在获能条件下孵育共培养物时,观察到更多存活(<0.05)、获能(<0.05)、具有进行性运动能力(<0.05)且顶体完整的精子群体(<0.05),这表明体外从OECs释放的精子群体具有潜在受精能力。改进冷冻精子的处理和选择将提高ARTs的效率,从而能够使用更高质量的精子群体。