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Protective roles of seminal plasma exosomes and microvesicles during human sperm cryopreservation

微泡 精子 低温保存 精子活力 精液 男科 运动性 生物 细胞生物学 化学 分子生物学 生物化学 胚胎 解剖 医学 小RNA 基因
作者
Forough Mahdavinezhad,Mohammad Ali Sadighi Gilani,Roghaye Gharaei,Zhaleh Ashrafnezhad,Jamal Valipour,Maryam Shabani Nashtaei,Fardin Amidi
出处
期刊:Reproductive Biomedicine Online [Elsevier BV]
卷期号:45 (2): 341-353 被引量:13
标识
DOI:10.1016/j.rbmo.2022.03.033
摘要

Abstract

Research question

Do seminal plasma microvesicles and exosomes, as two subtypes of extracellular vesicles, exert cryoprotective properties in sperm cryopreservation?

Design

Microvesicles and exosomes isolated from normozoospermic semen samples (n = 10) by serial ultracentrifugation were determined using scanning electron microscopy, dynamic light scattering and western blot analysis. The interactions between extracellular vesicles and spermatozoa were detected using Dil labelling. Purified spermatozoa from different normozoospermic samples (n = 25) were then treated individually with exosomes or microvesicles for 1 h and subsequently cryopreserved. The effects of extracellular vesicles during cryopreservation were investigated by determining post-thaw sperm motility, morphology, viability, reactive oxygen species (ROS) generation, lipid peroxidation, total antioxidant capacity (TAC), mitochondrial membrane potential (MMP), DNA integrity, and apoptosis rate.

Results

Microvesicles and exosomes displayed a round-shape morphology, with about 70% of exosomes ranging from 43–144 nm, microvesicles ranging from 144.5–486 nm and both expressed tetraspanin markers. Fluorescence microscopy showed that exosomes and microvesicles absorbed mainly in the sperm head and less frequently in the neck and tail. The post-thawing results indicated that the diluent with exosomes or microvesicles had improved sperm motility (P = 0.007), morphology (P < 0.001) and viability (P < 0.001) compared with untreated samples. The ROS levels decreased significantly (P = 0.001), with a consequent decrease in DNA damage (P = 0.001). The TAC activity (P = 0.001) and MMP levels (P = 0.001) were also significantly improved; levels of malondialdehyde (MDA) (P = 0.62) and apoptosis rate (P = 1.000) remained unchanged.

Conclusion

Seminal plasma microvesicles and exosomes could protect spermatozoa from cryopreservation chilling injuries.
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