Measurable Cytokine Concentrations in Pig Seminal Plasma Are Modified by Semen Handling and Storage

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-...

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Bibliographic Details
Published in:Biology (Basel, Switzerland) Switzerland), 2020-09, Vol.9 (9), p.276
Main Authors: Padilla, Lorena, Barranco, Isabel, Parrilla, Inmaculada, Lucas, Xiomara, Rodriguez-Martinez, Heriberto, Roca, Jordi
Format: Article
Language:English
Subjects:
Biomarkers
Cytokines
Ejaculation
Embryos
Experiments
Fluids
Granulocyte-macrophage colony-stimulating factor
IL-1β
Infertility
Interleukin 1 receptor antagonist
Interleukin 1 receptors
Interleukin 10
Interleukin 12
Interleukin 18
Interleukin 2
Interleukin 4
Interleukin 6
Interleukin 8
Medical laboratories
Physiological aspects
pig
Plasma
Semen
seminal plasma
Sperm
Storage
Storage conditions
Swine
Temperature
Tumor necrosis factor-α
γ-Interferon
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Summary: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.
ISSN:2079-7737
2079-7737
DOI:10.3390/biology9090276