Isolation of CD63-positive epididymosomes from human semen and its application in improving sperm function

IF 15.5 1区 医学 Q1 CELL BIOLOGY
Jingwen Luo, Shiqing Zhu, Yafei Kang, Xinyu Liu, Xia Tan, Jieyi Zhao, Xiaofang Ding, Honggang Li
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Abstract

Extracellular vesicles (EVs) are highly heterogeneous, and different EV subpopulations from various origins mediate different biological effects. The separation of different subpopulations of EVs from mixtures is critical but challenging. Epididymosomes are secreted by the epididymal epithelium and play a key role in sperm maturation and function. However, limited access to human epididymal tissue and epididymal fluid has hampered further study of epididymosomes and their potential clinical applications. Here, we established a novel strategy based on flow cytometry sorting to isolate human CD63-positive epididymosomes from ejaculate. We identified CD52, a membrane-located protein expressed exclusively in the epididymis, as the sorting marker for human epididymosomes. Then, CD63-positive epididymosomes were isolated from human semen using a flow cytometry sorting instrument and concentrated. Additionally, we observed that isolated CD63-positive epididymosomes improved sperm function more than other CD63-positive seminal EV subpopulations did, demonstrating the successful isolation of a subpopulation of epididymosomes from human semen and their potential application in the clinic.

从人类精液中分离 CD63 阳性附睾体及其在改善精子功能中的应用
细胞外囊泡(EVs)具有高度异质性,来自不同来源的不同 EVs 亚群会介导不同的生物效应。从混合物中分离出不同亚群的EVs至关重要,但也极具挑战性。附睾小体由附睾上皮分泌,在精子成熟和功能方面发挥着关键作用。然而,由于获取人类附睾组织和附睾液的途径有限,阻碍了对附睾小体及其潜在临床应用的进一步研究。在这里,我们建立了一种基于流式细胞术分选的新策略,从射精中分离出 CD63 阳性的人类附睾体。我们确定了 CD52(一种只在附睾中表达的膜定位蛋白)作为人类附睾体的分选标记。然后,我们使用流式细胞术分选仪从人类精液中分离出 CD63 阳性附睾体,并将其浓缩。此外,我们还观察到分离出的 CD63 阳性附睾小体比其他 CD63 阳性精液 EV 亚群更能改善精子功能,这表明我们成功地从人类精液中分离出了附睾小体亚群,并将其应用于临床。
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来源期刊
Journal of Extracellular Vesicles
Journal of Extracellular Vesicles Biochemistry, Genetics and Molecular Biology-Cell Biology
CiteScore
27.30
自引率
4.40%
发文量
115
审稿时长
12 weeks
期刊介绍: The Journal of Extracellular Vesicles is an open access research publication that focuses on extracellular vesicles, including microvesicles, exosomes, ectosomes, and apoptotic bodies. It serves as the official journal of the International Society for Extracellular Vesicles and aims to facilitate the exchange of data, ideas, and information pertaining to the chemistry, biology, and applications of extracellular vesicles. The journal covers various aspects such as the cellular and molecular mechanisms of extracellular vesicles biogenesis, technological advancements in their isolation, quantification, and characterization, the role and function of extracellular vesicles in biology, stem cell-derived extracellular vesicles and their biology, as well as the application of extracellular vesicles for pharmacological, immunological, or genetic therapies. The Journal of Extracellular Vesicles is widely recognized and indexed by numerous services, including Biological Abstracts, BIOSIS Previews, Chemical Abstracts Service (CAS), Current Contents/Life Sciences, Directory of Open Access Journals (DOAJ), Journal Citation Reports/Science Edition, Google Scholar, ProQuest Natural Science Collection, ProQuest SciTech Collection, SciTech Premium Collection, PubMed Central/PubMed, Science Citation Index Expanded, ScienceOpen, and Scopus.
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