The Secretome From Human Amniotic Membrane-Derived Mesenchymal Stem Cells Alleviate Oxidative Damage in Bovine Sperm Publisher Pubmed



Amirjannati N ; Barfar M ; Pournourali M ; Khalili N ; Tajik P ; Shams R ; Jahandideh A ; Goli LB ; Hashemian SJ ; Shirazi A ; Golshahi H
Source: Reproductive Biology Published:2026

Abstract

Bovine spermatozoa are highly vulnerable to oxidative stress (OS), which can impair motility, viability, membrane integrity, and DNA integrity. Human amniotic membrane-derived mesenchymal stem cells (AM-MSCs) secrete paracrine factors with antioxidant properties, making their secretome (AM-MSC-Sec) a potential tool for protecting sperm function. This study evaluated the protective effects of AM-MSC-Sec on bovine sperm under OS. Frozen Holstein bull semen was thawed, purified using a two-layer density gradient, and divided into four groups: control (no treatment), H₂O₂ (10 μM), H₂O₂ + 20 % AM-MSC-Sec, and H₂O₂ + 20 % DMEM/F12. After 2 h at 38.5°C and 5 % CO₂, sperm quality parameters—including motility, progressive motility, viability, plasma membrane integrity, acrosome reaction, DNA fragmentation, malondialdehyde (MDA), and total antioxidant capacity (TAC)—were assessed. AM-MSC-Sec significantly improved the percentage of total and progressive motility, enhanced viability and membrane integrity, and decreased DNA fragmentation compared with control and DMEM groups. The percentage of acrosome-reacted sperm increased, and TAC levels were elevated, indicating improved antioxidant defense. DMEM/F12 alone did not show a protective effect. MDA levels increased non-significantly under stress in the control group; however, the difference was not statistically significant. These findings suggest that AM-MSC-Sec could mitigate oxidative damage and preserve multiple functional parameters in bovine sperm. AM-MSC-Sec may serve as a promising therapeutic agent for maintaining sperm quality, with potential applications in bovine reproductive technologies. Copyright © 2025. Published by Elsevier B.V.