Spermatozoa and seminal plasma small extracellular vesicles miRNAs as biomarkers of boar semen cryotolerance

Freeze boar semen is still the biggest challenge for the swine industry due to the high cold shock sensitivity of boar sperm cells and the variance of post-thaw results among individuals and ejaculates from the same boar. To solve this problem, we investigate if miRNAs present in sperm cells and small extracellular vesicles (EVs) from seminal plasma of raw boar ejaculates can predict high-quality ejaculates after underwent the freeze-thaw process. For this, we obtained miRNAs samples of sperm cells and EVs from raw seminal plasma from 27 ejaculates before the cryopreservation process. Two groups with different freezability considering the analysis post-thaw of structure and sperm functionality were formed: High freezability (HF; n = 04) and low freezability (LF; n = 04). That done, we investigated the miRNAs profile of sperm cells and EVs from seminal plasma in both groups. Three miRNAs were differently abundant in LF ejaculates, being the ssc-miR-503 found in higher levels in sperm cells (P