Journal of Extracellular Vesicles最新文献

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A novel multiplexed immunoassay for surface-exposed proteins in plasma extracellular vesicles 血浆细胞外囊泡中表面暴露蛋白质的新型多重免疫测定法
IF 15.5 1区 医学
Journal of Extracellular Vesicles Pub Date : 2024-11-05 DOI: 10.1002/jev2.70007
Emma Tordoff, Jillian Allen, Katya Elgart, Ahmed Elsherbini, Vrinda Kalia, Haotian Wu, Erden Eren, Dimitrios Kapogiannis, Olesia Gololobova, Kenneth Witwer, Olga Volpert, Erez Eitan
{"title":"A novel multiplexed immunoassay for surface-exposed proteins in plasma extracellular vesicles","authors":"Emma Tordoff,&nbsp;Jillian Allen,&nbsp;Katya Elgart,&nbsp;Ahmed Elsherbini,&nbsp;Vrinda Kalia,&nbsp;Haotian Wu,&nbsp;Erden Eren,&nbsp;Dimitrios Kapogiannis,&nbsp;Olesia Gololobova,&nbsp;Kenneth Witwer,&nbsp;Olga Volpert,&nbsp;Erez Eitan","doi":"10.1002/jev2.70007","DOIUrl":"https://doi.org/10.1002/jev2.70007","url":null,"abstract":"<p>Small membranous extracellular vesicles (EV) incorporate proteins and nucleic acids from the parent cell. Proteins exposed on EV surface are dictated by cellular origin and biogenesis pathway. To better understand the EV origin and function, it is important to develop methods that reveal surface protein composition of heterogeneous EV populations in culture supernatants and in biofluids. Tetraspanins CD9, CD63, and CD81 are common and abundant EV markers. However, their relative enrichment (profile) on EVs of specific cellular origins is not fully elucidated. We introduce LuminEV, a novel version of the Luminex assay for the multiplexed analysis of EV surface proteins. Optimized LuminEV reagents enable direct, specific, and sensitive measurements of EV markers in biofluids and in culture supernatants, bypassing EV isolation step. LuminEV assay for CD9, CD63, and CD81 was validated by comparing simplex and multiplex measurements, establishing linearity, spike-in recovery, inter- and intra-assay precision, and reproducibility between operators. LuminEV measurements of CD9, CD63, and CD81 in conditioned media from 15 cell lines revealed strong variations between cell types and showed high sensitivity, which enabled EV detection without prior concentration. Using tetraspanin levels as a readout, we noted suppression and induction of EV release from the cultured cells by GW6869 and monensin. Measurement of EV CD9, CD63, and CD81 in blood plasma from 70 disease-free donors showed respective abundance of 72, 16, and 12%. CD63 displayed weak, albeit significant, negative correlation with age and was slightly lower in female samples. The assay was then used to detect cell type-specific EV surface markers, including CD235a (erythrocytes), GAP43 (neurons), and CD68 (macrophages), and to detect differences in tetraspanin profiles between healthy and diseased donors. In summary, LuminEV offers robust and sensitive approach for multiplexed assessment of EV surface proteins, to facilitate the research into EV biology, biomarker, and therapeutic applications.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":null,"pages":null},"PeriodicalIF":15.5,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70007","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142579595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Correction to MAP kinase kinase 1 (MEK1) within extracellular vesicles inhibits tumour growth by promoting anti-tumour immunity 校正细胞外囊泡内的 MAP 激酶 1 (MEK1) 可通过促进抗肿瘤免疫抑制肿瘤生长
IF 15.5 1区 医学
Journal of Extracellular Vesicles Pub Date : 2024-11-05 DOI: 10.1002/jev2.70010
{"title":"Correction to MAP kinase kinase 1 (MEK1) within extracellular vesicles inhibits tumour growth by promoting anti-tumour immunity","authors":"","doi":"10.1002/jev2.70010","DOIUrl":"https://doi.org/10.1002/jev2.70010","url":null,"abstract":"<p>Searles, S. C., Chen, W.-S., Yee, J. D., Lee, P., Lee, C. K., Caron, C., Mousovich-Neto, F., Matei, I., Lyden, D., &amp; Bui, J. D. (2024). MAP kinase kinase 1 (MEK1) within extracellular vesicles inhibits tumour growth by promoting anti-tumour immunity. <i>Journal of Extracellular Vesicles</i>, 13, e12515. https://doi.org/10.1002/jev2.12515</p><p>In the originally-published article, author Felippe Mousovich-Neto's name was incorrectly given as Felippe Neto. The online version of the article has been corrected.</p><p>We apologize for this error.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":null,"pages":null},"PeriodicalIF":15.5,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70010","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142579582","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A switch from lysosomal degradation to secretory autophagy initiates osteogenic bone metastasis in prostate cancer 从溶酶体降解到分泌型自噬的转换启动了前列腺癌的成骨性骨转移。
IF 15.5 1区 医学
Journal of Extracellular Vesicles Pub Date : 2024-11-05 DOI: 10.1002/jev2.70002
Xiaoyu Wei, Mengmeng Liang, Min Deng, Ji Zheng, Fei Luo, Qinyu Ma
{"title":"A switch from lysosomal degradation to secretory autophagy initiates osteogenic bone metastasis in prostate cancer","authors":"Xiaoyu Wei,&nbsp;Mengmeng Liang,&nbsp;Min Deng,&nbsp;Ji Zheng,&nbsp;Fei Luo,&nbsp;Qinyu Ma","doi":"10.1002/jev2.70002","DOIUrl":"10.1002/jev2.70002","url":null,"abstract":"<p>The identification of both autophagy-related material degradation and unconventional secretion has paved the way for significant breakthroughs linking autophagy to a plethora of physiological processes and disease conditions. However, the mechanisms that coordinate these two pathways remain elusive. Here, we demonstrate that a switch from the lysosomal degradation to a secretory autophagy pathway is governed by protein tyrosine phosphatase 1B (PTP1B, encoded by <i>PTPN1</i>). Dephosphorylation at two tyrosine residues of syntaxin17 (STX17) by PTP1B reduces autophagosome-lysosome fusion while switching the cells to a secretory autophagy pathway. Both PTP1B overexpression and tumour-derived extracellular vesicles (EVs) can activate the secretory autophagy pathway in osteoblasts. Moreover, we demonstrate that osteoblastic LC3+ EVs, generated via the secretory autophagy pathway, are the primary contributor to tumour-associated bone remodelling in prostate cancer. Depletion of tumour-derived EVs secretion or genetic ablation of osteoblastic PTP1B rescues aberrant bone remodelling and lesions, highlighting the relevance between LC3+ EVs and the formation of bone metastatic niche. Our results reveal the significance of tumour-regulated PTP1B in the fate decision of autophagosomes, and propose a role ofLC3+ EVs in shaping the bone metastatic niche.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":null,"pages":null},"PeriodicalIF":15.5,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70002","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142576330","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Extracellular vesicles from human-induced pluripotent stem cell-derived neural stem cells alleviate proinflammatory cascades within disease-associated microglia in Alzheimer's disease 来自人类诱导多能干细胞衍生神经干细胞的细胞外囊泡能减轻阿尔茨海默病中与疾病相关的小胶质细胞内的促炎级联反应
IF 15.5 1区 医学
Journal of Extracellular Vesicles Pub Date : 2024-11-05 DOI: 10.1002/jev2.12519
Leelavathi N. Madhu, Maheedhar Kodali, Raghavendra Upadhya, Shama Rao, Yogish Somayaji, Sahithi Attaluri, Bing Shuai, Maha Kirmani, Shreyan Gupta, Nathaniel Maness, Xiaolan Rao, James J. Cai, Ashok K. Shetty
{"title":"Extracellular vesicles from human-induced pluripotent stem cell-derived neural stem cells alleviate proinflammatory cascades within disease-associated microglia in Alzheimer's disease","authors":"Leelavathi N. Madhu,&nbsp;Maheedhar Kodali,&nbsp;Raghavendra Upadhya,&nbsp;Shama Rao,&nbsp;Yogish Somayaji,&nbsp;Sahithi Attaluri,&nbsp;Bing Shuai,&nbsp;Maha Kirmani,&nbsp;Shreyan Gupta,&nbsp;Nathaniel Maness,&nbsp;Xiaolan Rao,&nbsp;James J. Cai,&nbsp;Ashok K. Shetty","doi":"10.1002/jev2.12519","DOIUrl":"https://doi.org/10.1002/jev2.12519","url":null,"abstract":"<p>As current treatments for Alzheimer's disease (AD) lack disease-modifying interventions, novel therapies capable of restraining AD progression and maintaining better brain function have great significance. Anti-inflammatory extracellular vesicles (EVs) derived from human induced pluripotent stem cell (hiPSC)-derived neural stem cells (NSCs) hold promise as a disease-modifying biologic for AD. This study directly addressed this issue by examining the effects of intranasal (IN) administrations of hiPSC-NSC-EVs in 3-month-old 5xFAD mice. IN administered hiPSC-NSC-EVs incorporated into microglia, including plaque-associated microglia, and encountered astrocyte soma and processes in the brain. Single-cell RNA sequencing revealed transcriptomic changes indicative of diminished activation of microglia and astrocytes. Multiple genes linked to disease-associated microglia, NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3)-inflammasome and interferon-1 (IFN-1) signalling displayed reduced expression in microglia. Adding hiPSC-NSC-EVs to cultured human microglia challenged with amyloid-beta oligomers resulted in similar effects. Astrocytes also displayed reduced expression of genes linked to IFN-1 and interleukin-6 signalling. Furthermore, the modulatory effects of hiPSC-NSC-EVs on microglia in the hippocampus persisted 2 months post-EV treatment without impacting their phagocytosis function. Such effects were evidenced by reductions in microglial clusters and inflammasome complexes, concentrations of mediators, and end products of NLRP3 inflammasome activation, the expression of genes and/or proteins involved in the activation of p38/mitogen-activated protein kinase and IFN-1 signalling, and unaltered phagocytosis function. The extent of astrocyte hypertrophy, amyloid-beta plaques, and p-tau were also reduced in the hippocampus. Such modulatory effects of hiPSC-NSC-EVs also led to better cognitive and mood function. Thus, early hiPSC-NSC-EV intervention in AD can maintain better brain function by reducing adverse neuroinflammatory signalling cascades, amyloid-beta plaque load, and p-tau. These results reflect the first demonstration of the efficacy of hiPSC-NSC-EVs to restrain neuroinflammatory signalling cascades in an AD model by inducing transcriptomic changes in activated microglia and reactive astrocytes.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":null,"pages":null},"PeriodicalIF":15.5,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.12519","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142579729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Real-time monitoring of small extracellular vesicles (sEVs) by in vivo flow cytometry 利用体内流式细胞仪实时监测细胞外小泡(sEVs)。
IF 15.5 1区 医学
Journal of Extracellular Vesicles Pub Date : 2024-10-23 DOI: 10.1002/jev2.70003
Fuli Zhang, Xin Lu, Xi Zhu, Ziwen Yu, Weiliang Xia, Xunbin Wei
{"title":"Real-time monitoring of small extracellular vesicles (sEVs) by in vivo flow cytometry","authors":"Fuli Zhang,&nbsp;Xin Lu,&nbsp;Xi Zhu,&nbsp;Ziwen Yu,&nbsp;Weiliang Xia,&nbsp;Xunbin Wei","doi":"10.1002/jev2.70003","DOIUrl":"10.1002/jev2.70003","url":null,"abstract":"<p>Extracellular vesicles (EVs) are vesicular structures comprised of a bilayer lipid membrane, released by living cells. There is a growing body of evidence for their functionality, indicating that small EVs (sEVs) can mediate specific forms of intercellular communication. The future applications of sEVs hold great promise, not only as diagnostic markers but also as therapeutic agents. However, the greatest difficulty in the clinical translation of sEVs is that they are currently poorly understood, especially concerning their in vivo behaviour. In this study, we provide a novel method for monitoring sEVs in blood circulation based on in vivo flow cytometry (IVFC). We have demonstrated that the height of the IVFC signal baseline is proportional to the concentration of sEVs. Moreover, we have found out that the peaks in the IVFC signal are generated by the aggregation or cellular uptake of sEVs. In vivo monitoring of sEVs clearance from the blood indicates that PEGylated sEVs have a longer residence time and exhibit less aggregation in circulation compared to non-PEGylated sEVs. These studies reveal that IVFC enables real-time in vivo monitoring of circulating sEVs, which can provide valuable insights into the pharmacokinetics and cellular targeting capabilities of sEVs.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":null,"pages":null},"PeriodicalIF":15.5,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11497658/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142501740","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Separation of small extracellular vesicles (sEV) from human blood by Superose 6 size exclusion chromatography 用 Superose 6 尺寸排阻色谱法分离人血中的小细胞外囊泡 (sEV)。
IF 15.5 1区 医学
Journal of Extracellular Vesicles Pub Date : 2024-10-23 DOI: 10.1002/jev2.70008
Jerome Nouvel, Gonzalo Bustos Quevedo, Tony Prinz, Ramsha Masood, George Daaboul, Tanja Gainey-Schleicher, Uwe Wittel, Sophia Chikhladze, Bence Melykuti, Martin Helmstaedter, Karl Winkler, Irina Nazarenko, Gerhard Pütz
{"title":"Separation of small extracellular vesicles (sEV) from human blood by Superose 6 size exclusion chromatography","authors":"Jerome Nouvel,&nbsp;Gonzalo Bustos Quevedo,&nbsp;Tony Prinz,&nbsp;Ramsha Masood,&nbsp;George Daaboul,&nbsp;Tanja Gainey-Schleicher,&nbsp;Uwe Wittel,&nbsp;Sophia Chikhladze,&nbsp;Bence Melykuti,&nbsp;Martin Helmstaedter,&nbsp;Karl Winkler,&nbsp;Irina Nazarenko,&nbsp;Gerhard Pütz","doi":"10.1002/jev2.70008","DOIUrl":"10.1002/jev2.70008","url":null,"abstract":"<p>Extracellular vesicles (EVs) are valuable targets for liquid biopsy. However, attempts to introduce EV-based biomarkers into clinical practice have not been successful to the extent expected. One of the reasons for this failure is the lack of reliable methods for EV baseline purification from complex biofluids, such as cell-free plasma or serum. Because available one-step approaches for EV isolation are insufficient to purify EVs, the majority of studies on clinical samples were performed either on a mixture of EVs and lipoproteins, whilst the real number of EVs and their individual specific biomarker content remained elusive, or on a low number of samples of sufficient volume to allow elaborate 2-step EV separation by size and density, resulting in a high purity but utmost low recovery. Here we introduce Fast Protein Liquid Chromatography (FPLC) using Superose 6 as a matrix to obtain small EVs from biofluids that are almost free of soluble proteins and lipoproteins. Along with the estimation of a realistic number of small EVs in human samples, we show temporal resolution of the effect of the duration of postprandial phase on the proportion of lipoproteins in purified EVs, suggesting acceptable time frames additionally to the recommendation to use fasting samples for human studies. Furthermore, we assessed a potential value of pure EVs for liquid biopsy, exemplarily examining EV- and tumour-biomarkers in pure FPLC-derived fractions isolated from the serum of patients with pancreatic cancer. Consistent among different techniques, showed the presence of diseases-associated biomarkers in pure EVs, supporting the feasibility of using single-vesicle analysis for liquid biopsy.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":null,"pages":null},"PeriodicalIF":15.5,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11497763/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142501741","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Bacterial extracellular vesicles as intranasal postbiotics: Detailed characterization and interaction with airway cells 作为鼻内后生物制剂的细菌胞外囊泡:详细表征及与气道细胞的相互作用
IF 15.5 1区 医学
Journal of Extracellular Vesicles Pub Date : 2024-10-21 DOI: 10.1002/jev2.70004
Agnieszka Razim, Agnieszka Zabłocka, Anna Schmid, Michael Thaler, Viktor Černý, Tamara Weinmayer, Bradley Whitehead, Anke Martens, Magdalena Skalska, Mattia Morandi, Katy Schmidt, Magdalena E. Wysmołek, Akos Végvári, Dagmar Srutkova, Martin Schwarzer, Lukas Neuninger, Peter Nejsum, Jiri Hrdý, Johan Palmfeldt, Marco Brucale, Francesco Valle, Sabina Górska, Lukas Wisgrill, Aleksandra Inic-Kanada, Ursula Wiedermann, Irma Schabussova
{"title":"Bacterial extracellular vesicles as intranasal postbiotics: Detailed characterization and interaction with airway cells","authors":"Agnieszka Razim,&nbsp;Agnieszka Zabłocka,&nbsp;Anna Schmid,&nbsp;Michael Thaler,&nbsp;Viktor Černý,&nbsp;Tamara Weinmayer,&nbsp;Bradley Whitehead,&nbsp;Anke Martens,&nbsp;Magdalena Skalska,&nbsp;Mattia Morandi,&nbsp;Katy Schmidt,&nbsp;Magdalena E. Wysmołek,&nbsp;Akos Végvári,&nbsp;Dagmar Srutkova,&nbsp;Martin Schwarzer,&nbsp;Lukas Neuninger,&nbsp;Peter Nejsum,&nbsp;Jiri Hrdý,&nbsp;Johan Palmfeldt,&nbsp;Marco Brucale,&nbsp;Francesco Valle,&nbsp;Sabina Górska,&nbsp;Lukas Wisgrill,&nbsp;Aleksandra Inic-Kanada,&nbsp;Ursula Wiedermann,&nbsp;Irma Schabussova","doi":"10.1002/jev2.70004","DOIUrl":"https://doi.org/10.1002/jev2.70004","url":null,"abstract":"<p><i>Escherichia coli</i> A0 34/86 (EcO83) is a probiotic strain used in newborns to prevent nosocomial infections and diarrhoea. This bacterium stimulates both pro- and anti-inflammatory cytokine production and its intranasal administration reduces allergic airway inflammation in mice. Despite its benefits, there are concerns about the use of live probiotic bacteria due to potential systemic infections and gene transfer. Extracellular vesicles (EVs) derived from EcO83 (EcO83-EVs) might offer a safer alternative to live bacteria. This study characterizes EcO83-EVs and investigates their interaction with host cells, highlighting their potential as postbiotic therapeutics. EcO83-EVs were isolated, purified, and characterised following the Minimal Information of Studies of Extracellular Vesicles (MISEV) guidelines. Ex vivo studies conducted in human nasal epithelial cells showed that EcO83-EVs increased the expression of proteins linked to oxidative stress and inflammation, indicating an effective interaction between EVs and the host cells. Further in vivo studies in mice demonstrated that EcO83-EVs interact with nasal-associated lymphoid tissue, are internalised by airway macrophages, and stimulate neutrophil recruitment in the lung. Mechanistically, EcO83-EVs activate the NF-κΒ signalling pathway, resulting in the nitric oxide production. EcO83-EVs demonstrate significant potential as a postbiotic alternative to live bacteria, offering a safer option for therapeutic applications. Further research is required to explore their clinical use, particularly in mucosal vaccination and targeted immunotherapy strategies.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":null,"pages":null},"PeriodicalIF":15.5,"publicationDate":"2024-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142451785","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Isolation of CD63-positive epididymosomes from human semen and its application in improving sperm function 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
IF 15.5 1区 医学
Journal of Extracellular Vesicles Pub Date : 2024-10-17 DOI: 10.1002/jev2.70006
Jingwen Luo, Shiqing Zhu, Yafei Kang, Xinyu Liu, Xia Tan, Jieyi Zhao, Xiaofang Ding, Honggang Li
{"title":"Isolation of CD63-positive epididymosomes from human semen and its application in improving sperm function","authors":"Jingwen Luo,&nbsp;Shiqing Zhu,&nbsp;Yafei Kang,&nbsp;Xinyu Liu,&nbsp;Xia Tan,&nbsp;Jieyi Zhao,&nbsp;Xiaofang Ding,&nbsp;Honggang Li","doi":"10.1002/jev2.70006","DOIUrl":"https://doi.org/10.1002/jev2.70006","url":null,"abstract":"<p>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.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":null,"pages":null},"PeriodicalIF":15.5,"publicationDate":"2024-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70006","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142447775","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Size photometry and fluorescence imaging of immobilized immersed extracellular vesicles 固定沉浸式细胞外囊泡的尺寸光度测量和荧光成像
IF 15.5 1区 医学
Journal of Extracellular Vesicles Pub Date : 2024-10-14 DOI: 10.1002/jev2.12512
Andreas Wallucks, Philippe DeCorwin-Martin, Molly L. Shen, Andy Ng, David Juncker
{"title":"Size photometry and fluorescence imaging of immobilized immersed extracellular vesicles","authors":"Andreas Wallucks,&nbsp;Philippe DeCorwin-Martin,&nbsp;Molly L. Shen,&nbsp;Andy Ng,&nbsp;David Juncker","doi":"10.1002/jev2.12512","DOIUrl":"https://doi.org/10.1002/jev2.12512","url":null,"abstract":"<p>Immunofluorescence analysis of individual extracellular vesicles (EVs) in common fluorescence microscopes is gaining popularity due to its accessibility and high fluorescence sensitivity; however, EV number and size are only measurable using fluorescent stains requiring extensive sample manipulations. Here we introduce highly sensitive label-free EV size photometry (SP) based on interferometric scattering (iSCAT) imaging of immersed EVs immobilized on a glass coverslip. We implement SP on a common inverted epifluorescence microscope with LED illumination and a simple 50:50 beamsplitter, permitting seamless integration of SP with fluorescence imaging (SPFI). We present a high-throughput SPFI workflow recording &gt;10,000 EVs in 7 min over ten 88 × 88 µm<sup>2</sup> fields of view, pre- and post-incubation imaging to suppress background, along with automated image alignment, aberration correction, spot detection and EV sizing. We achieve an EV sizing range from 37 to ∼220 nm in diameter with a dual 440 and 740 nm SP illumination scheme, and suggest that this range can be extended by more advanced image analysis or additional hardware customization. We benchmark SP to flow cytometry using calibrated silica nanoparticles and demonstrate superior, label-free sensitivity. We showcase SPFI's potential for EV analysis by experimentally distinguishing surface and volumetric EV dyes, observing the deformation of EVs adsorbed to a surface, and by uncovering distinct subpopulations in &lt;100 nm-in-diameter EVs with fluorescently tagged membrane proteins.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":null,"pages":null},"PeriodicalIF":15.5,"publicationDate":"2024-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.12512","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142435604","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Low fluid shear stress stimulates the uptake of noxious endothelial extracellular vesicles via MCAM and PECAM-1 cell adhesion molecules 低流体剪切应力通过 MCAM 和 PECAM-1 细胞粘附分子刺激有害内皮细胞外囊泡的吸收
IF 15.5 1区 医学
Journal of Extracellular Vesicles Pub Date : 2024-10-14 DOI: 10.1002/jev2.12414
Pierre-Michaël Coly, Shruti Chatterjee, Fariza Mezine, Christelle El Jekmek, Cécile Devue, Thomas Nipoti, Stephane Mazlan, Maribel Lara Corona, Florent Dingli, Damarys Loew, Guillaume van Niel, Xavier Loyer, Chantal M. Boulanger
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