: Seminal Extracellular Vesicles – A New Frontier in Biomarker Discovery Through Protocol Optimization

ABSTRACT: : Seminal extracellular vesicles (EVs) hold a promise as a source of biomarkers for urogenital diseases. Unlocking their potential in clinical research requires further investigation to optimize the isolation protocols of these vesicles from semen. This study compares different isolation techniques of small seminal EVs, focusing on ultracentrifugation (UC) and size-exclusion chromatography (SEC). The protein content of the EV preparations was then analyzed by mass spectrometry . Our results show that EV samples prepared by SEC or by UC were enriched in different proteins. EV-UC preparations contained more typical EV-related molecules compared to EV-SEC preparations. On the other hand, ribosomal proteins and ER-related proteins were enriched in SEC-EV samples compared to UC-EV samples. Furthermore, expected soluble proteins in semen samples were more abundant in EVs isolated by SEC. These results suggest that seminal EVs originate from various mechanisms and or cellular sites. Moreover, UC was more efficient in removing soluble proteins from EV samples than SEC. Based on these results, the EV isolation method should be carefully considered in relation to the study purpose.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Ingrid Maria Erika Ekman Stensland  

LAB HEAD: Tuula Anneli Nyman

PROVIDER: PXD069636 | Pride | 2026-10-26

REPOSITORIES: Pride