Measurable Cytokine Concentrations in Pig Seminal Plasma Are Modified by Semen Handling and Storage
Open Access
- 7 September 2020
- Vol. 9 (9), 276
- https://doi.org/10.3390/biology9090276
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.Keywords
Funding Information
- European Regional Development Fund (AGL2015-69738-R)
This publication has 53 references indexed in Scilit:
- Multiplex serum biomarker assessments: technical and biostatistical issuesJournal of Translational Medicine, 2011
- Prerequisites for cytokine measurements in clinical trials with multiplex immunoassaysBMC Immunology, 2009
- Seminal Plasma Cytokines and Chemokines in Prostate Inflammation: Interleukin 8 as a Predictive Biomarker in Chronic Prostatitis/Chronic Pelvic Pain Syndrome and Benign Prostatic HyperplasiaEuropean Urology, 2006
- Impact of storage prior to cryopreservation on plasma membrane function and fertility of boar spermTheriogenology, 2005
- Regulation of sperm function by reactive oxygen speciesHuman Reproduction Update, 2004
- Multiplex bead array assays for detection of soluble cytokines: Comparisons of sensitivity and quantitative values among kits from multiple manufacturersCytometry Part B: Clinical Cytometry, 2004
- The fecundity of porcine semen stored for 2 to 6 days in androhep® and X-cell™ extendersTheriogenology, 1999
- Influence of collection and separation of blood samples on plasma IL-1, IL-6 and TNF-α concentrationsJournal of Immunological Methods, 1992