Abstract
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 (n = 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.
Highlights
Seminal plasma (SP) is a complex fluid, with biomolecules playing key roles in sperm function, fertilization and even embryo development and implantation [1,2]
The results showed that delaying seminal plasma (SP) harvesting by up to 24 h after ejaculate collection does not have a substantial impact on SP cytokine measurements
There are no ideal short- or long-term storage conditions for SP samples that allow for reliable measurement of the original levels of all cytokines of interest, at least not among those quantified in this study
Summary
Seminal plasma (SP) is a complex fluid, with biomolecules playing key roles in sperm function, fertilization and even embryo development and implantation [1,2]. SP components are involved in the regulation of the uterine immune environment facilitating the development and implantation of embryos [5]. SP components are currently being explored for potential biomarkers of male (in)fertility [6,7]. Among these biomolecules are cytokines, low-molecular-weight signaling proteins/peptides involved in intercellular communication, tissue homeostasis and the body’s immune response [8,9,10]. Functional changes in the male reproductive organs lead to differences in cytokine synthesis, differences in SP cytokine levels are mirrored in reproductive (dys)function [12,13,14]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
