Padilla Lorena, Barranco Isabel, Parrilla Inmaculada, Lucas Xiomara, Rodriguez-Martinez Heriberto, Roca Jordi
Department of Medicine and Animal Surgery, Veterinary Science, University of Murcia, 30100 Murcia, Spain.
IMIB-Arrixaca, Regional Campus of International Excellence, Campus Mare Nostrum, University of Murcia, 30100 Murcia, Spain.
Biology (Basel). 2020 Sep 7;9(9):276. doi: 10.3390/biology9090276.
Sample handling and storing are critical steps for the reliable measurement of circulating biomolecules in biological fluids. This study evaluates how cytokine measurements in pig seminal plasma (SP) vary depending on semen handling and SP storage. Thirteen cytokines (GM-CSF, IFNγ, IL-1α, IL-1β, IL-1ra, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-18 and TNFα) were measured using Luminex xMAP technology in individual seminal plasma (SP) samples ( = 62) from healthy breeding boars. Three separate experiments explored the delay (2 h and 24 h) in SP collection after ejaculation (Experiment 1) and SP storage, either short-term (5 °C, -20 °C and -80 °C for 72 h, Experiment 2) or long-term (at -20 °C and -80 °C for two months, Experiment 3), before analysis. Levels in fresh SP-samples were used as baseline control values. Delays in SP harvesting of up to 24 h did not substantially impact SP cytokine measurements. Some cytokines showed instability in stored SP samples, mainly in long-term storage. Ideally, cytokines in pig SP should be measured in fresh samples harvested within 24 h after ejaculation. If storage of SP is imperative, storage conditions should be adjusted for each cytokine.
样本处理和储存是可靠测量生物体液中循环生物分子的关键步骤。本研究评估了猪精液血浆（SP）中细胞因子的测量如何因精液处理和SP储存的不同而变化。使用Luminex xMAP技术对来自健康种公猪的个体精液血浆（SP）样本（n = 62）中的13种细胞因子（GM-CSF、IFNγ、IL-1α、IL-1β、IL-1ra、IL-2、IL-4、IL-6、IL-8、IL-10、IL-12、IL-18和TNFα）进行了测量。三个独立实验分别探讨了射精后SP采集的延迟时间（2小时和24小时，实验1）以及SP储存情况，包括短期储存（5°C、-20°C和-80°C下储存72小时，实验2）或长期储存（-20°C和-80°C下储存两个月，实验3），然后进行分析。新鲜SP样本中的水平用作基线对照值。SP采集延迟长达24小时对SP细胞因子测量没有实质性影响。一些细胞因子在储存的SP样本中表现出不稳定性，主要是在长期储存中。理想情况下，猪SP中的细胞因子应在射精后24小时内采集的新鲜样本中进行测量。如果必须储存SP，则应针对每种细胞因子调整储存条件。
