Journal of extracellular biology最新文献

{"title":"Profiling biomanufactured extracellular vesicles of human forebrain spheroids in a Vertical-Wheel Bioreactor","authors":"Chang Liu,&nbsp;Li Sun,&nbsp;Hannah Worden,&nbsp;Justice Ene,&nbsp;Olivia Z. Zeng,&nbsp;Jamini Bhagu,&nbsp;Samuel C. Grant,&nbsp;Xiaoping Bao,&nbsp;Sunghoon Jung,&nbsp;Yan Li","doi":"10.1002/jex2.70002","DOIUrl":"https://doi.org/10.1002/jex2.70002","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <p>Extracellular vesicles (EVs) secreted by human brain cells have great potential as cell-free therapies in various diseases, including stroke. However, because of the significant amount of EVs needed in preclinical and clinical trials, EV application is still challenging. Vertical-Wheel Bioreactors (VWBRs) have designed features that allow for scaling up the generation of human forebrain spheroid EVs under low shear stress.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <p>In this study, EV secretion by human forebrain spheroids derived from induced pluripotent stem cells as 3D aggregates and on Synthemax II microcarriers in VWBRs were investigated with static aggregate culture as a control. The spheroids were characterized by metabolite and transcriptome analysis. The isolated EVs were characterized by nanoparticle tracking analysis, electron microscopy, and Western blot. The EV cargo was analyzed using proteomics and miRNA sequencing. The in vitro functional assays of an oxygen and glucose-deprived stroke model were conducted. Proof of concept in vivo study was performed, too.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <p>Human forebrain spheroid differentiated on microcarriers showed a higher growth rate than 3D aggregates. Microcarrier culture had lower glucose consumption per million cells and lower glycolysis gene expression but higher EV biogenesis genes. EVs from the three culture conditions showed no differences in size, but the yields from high to low were microcarrier cultures, dynamic aggregates, and static aggregates. The cargo is enriched with proteins (proteomics) and miRNAs (miRNA-seq), promoting axon guidance, reducing apoptosis, scavenging reactive oxygen species, and regulating immune responses. Human forebrain spheroid EVs demonstrated the ability to improve recovery in an in vitro stroke model and in vivo.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <p>Human forebrain spheroid differentiation in VWBR significantly increased the EV yields (up to 240–750 fold) and EV biogenesis compared to static differentiation due to the dynamic microenvironment and metabolism change. The biomanufactured EVs from VWBRs have exosomal characteristics and more therapeutic cargo and are functional in in vitro assays, which paves the way for future in vivo stroke studies.</p>\u0000 </section>\u0000 </div>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"3 9","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70002","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142089842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineered extracellular vesicles coated with an antimicrobial peptide for advanced control of bacterial sepsis","authors":"Usri H. Ibrahim,&nbsp;Mohammed A. Gafar,&nbsp;Rene Khan,&nbsp;Abdelrahman Tageldin,&nbsp;Thirumala Govender,&nbsp;Irene Mackraj","doi":"10.1002/jex2.70000","DOIUrl":"https://doi.org/10.1002/jex2.70000","url":null,"abstract":"<p>Alarming sepsis-related mortality rates present significant challenges to healthcare services globally. Despite advances made in the field, there is still an urgent need to develop innovative approaches that could improve survival rates and reduce the overall cost of treatment for sepsis patients. Therefore, this study aimed to develop a novel multifunctional therapeutic agent for advanced control of bacterial sepsis. Extracellular vesicles (EVs) isolated from lipopolysaccharide (LPS) induced HepG2 (hepatocellular carcinoma cells) (iEV) displayed an average particle size of 171.63 ± 2.77 nm, a poly dispersion index (PDI) of 0.32 ± 0.0, and a zeta potential (ZP) of −11.87 ± 0.18 mV. Compared to HepG2 EV, LPS induction significantly increases the EV protein concentration, PDI and ZP, reduces the average size and promotes cell proliferation and cytoprotective effects of the isolated EVs (iEVs) against LPS-induced cytotoxicity. Coating of iEV with a cationic antimicrobial peptide (AMP) to form PC-iEV slightly changed their physical properties and shifted their surface charge toward neutral values. This modification improved the antibacterial activity (2-fold lower minimum bactericidal concentration [MBC] values) and biocompatibility of the conjugated peptide while maintaining iEV cytoprotective and anti-inflammatory activities. Our findings indicate the superior anti-inflammatory and antibacterial dual activity of PC-iEV against pathogens associated with sepsis.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"3 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70000","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142041570","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Orthogonal analysis reveals inconsistencies in cargo loading of extracellular vesicles","authors":"Neona M. Lowe,&nbsp;Rachel R. Mizenko,&nbsp;Bryan B. Nguyen,&nbsp;Kwan Lun Chiu,&nbsp;Vishalakshi Arun,&nbsp;Alyssa Panitch,&nbsp;Randy P. Carney","doi":"10.1002/jex2.70003","DOIUrl":"https://doi.org/10.1002/jex2.70003","url":null,"abstract":"<p>Since extracellular vesicles (EVs) have emerged as a promising drug delivery system, diverse methods have been used to load them with active pharmaceutical ingredients (API) in preclinical and clinical studies. However, there is yet to be an engineered EV formulation approved for human use, a barrier driven in part by the intrinsic heterogeneity of EVs. API loading is rarely assessed in the context of single vesicle measurements of physicochemical properties but is likely administered in a heterogeneous fashion to the detriment of a consistent product. Here, we applied a suite of single-particle resolution methods to determine the loading of rhodamine 6G (R6G) surrogate cargo mimicking hydrophilic small molecule drugs across four common API loading methods: sonication, electroporation, freeze-thaw cycling and passive incubation. Loading efficiencies and alterations in the physical properties of EVs were assessed, as well as co-localization with common EV-associated tetraspanins (i.e., CD63, CD81 and CD9) for insight into EV subpopulations. Sonication had the highest loading efficiency, yet significantly decreased particle yield, while electroporation led to the greatest number of loaded API particles, albeit at a lower efficiency. Moreover, results were often inconsistent between repeated runs within a given method, demonstrating the difficulty in developing a rigorous loading method that consistently loaded EVs across their heterogeneous subpopulations. This work highlights the significance of how chosen quantification metrics can impact apparent conclusions and the importance of single-particle characterization of EV loading.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"3 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142041569","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Growth phase matters: Boosting immunity via Lacticasebacillus-derived membrane vesicles and their interactions with TLR2 pathways","authors":"Miriam Sandanusova,&nbsp;Kristyna Turkova,&nbsp;Eva Pechackova,&nbsp;Jan Kotoucek,&nbsp;Pavel Roudnicky,&nbsp;Martin Sindelar,&nbsp;Lukas Kubala,&nbsp;Gabriela Ambrozova","doi":"10.1002/jex2.169","DOIUrl":"https://doi.org/10.1002/jex2.169","url":null,"abstract":"<p>Lipid bi-layered particles known as membrane vesicles (MVs), produced by Gram-positive bacteria are a communication tool throughout the entire bacterial growth. However, the MVs characteristics may vary across all stages of maternal culture growth, leading to inconsistencies in MVs research. This, in turn, hinders their employment as nanocarriers, vaccines and other medical applications. In this study, we aimed to comprehensively characterize MVs derived from <i>Lacticaseibacillus rhamnosus</i> CCM7091 isolated at different growth stages: early exponential (6 h, MV6), late exponential (12 h, MV12) and late stationary phase (48 h, MV48). We observed significant differences in protein content between MV6 and MV48 (data are available via ProteomeXchange with identifier PXD041580), likely contributing to their different immunomodulatory capacities. In vitro analysis demonstrated that MV48 uptake rate by epithelial Caco-2 cells is significantly higher and they stimulate an immune response in murine macrophages RAW 264.7 (elevated production of TNFα, IL-6, IL-10, NO). This correlated with increased expression of lipoteichoic acid (LTA) and enhanced TLR2 signalling in MV48, suggesting that LTA contributes to the immunomodulation. In conclusion, we showed that <i>Lacticaseibacillus rhamnosus</i> CCM7091-derived MVs from the late stationary phase boost the immune response the most effectively, which pre-destines them for therapeutical application as nanocarriers.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"3 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.169","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142041568","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular vesicles may provide an alternative detoxification pathway during skeletal muscle myoblast ageing","authors":"María Fernández-Rhodes,&nbsp;Emma Buchan,&nbsp;Stephanie D. Gagnon,&nbsp;Jiani Qian,&nbsp;Lee Gethings,&nbsp;Rebecca Lees,&nbsp;Ben Peacock,&nbsp;Andrew J. Capel,&nbsp;Neil R. W. Martin,&nbsp;Pola Goldberg Oppenheimer,&nbsp;Mark P. Lewis,&nbsp;Owen G. Davies","doi":"10.1002/jex2.171","DOIUrl":"10.1002/jex2.171","url":null,"abstract":"<p>Skeletal muscle (SM) acts as a secretory organ, capable of releasing myokines and extracellular vesicles (SM-EVs) that impact myogenesis and homeostasis. While age-related changes have been previously reported in murine SM-EVs, no study has comprehensively profiled SM-EV in human models. To this end, we provide the first comprehensive comparison of SM-EVs from young and old human primary skeletal muscle cells (HPMCs) to map changes associated with SM ageing. HPMCs, isolated from young (24 ± 1.7 years old) and older (69 ± 2.6 years old) participants, were immunomagnetically sorted based on the presence of the myogenic marker CD56 (N-CAM) and cultured as pure (100% CD56⁺) or mixed populations (MP: 90% CD56⁺). SM-EVs were isolated using an optimised protocol combining ultrafiltration and size exclusion chromatography (UF + SEC) and their biological content was extensively characterised using Raman spectroscopy (RS) and liquid chromatography mass spectrometry (LC-MS). Minimal variations in basic EV parameters (particle number, size, protein markers) were observed between young and old populations. However, biochemical fingerprinting by RS highlighted increased protein (amide I), lipid (phospholipids and phosphatidylcholine) and hypoxanthine signatures for older SM-EVs. Through LC-MS, we identified 84 shared proteins with functions principally related to cell homeostasis, muscle maintenance and transcriptional regulation. Significantly, SM-EVs from older participants were comparatively enriched in proteins involved in oxidative stress and DNA/RNA mutagenesis, such as E3 ubiquitin-protein ligase TTC3 (TTC3), little elongation complex subunit 1 (ICE1) and Acetyl-CoA carboxylase 1 (ACACA). These data suggest SM-EVs could provide an alternative pathway for homeostasis and detoxification during SM ageing.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"3 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11336379/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142019740","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Increased levels of circulating cell-free double-stranded nucleic acids in the plasma of glioblastoma patients","authors":"Elisabeth Rackles,&nbsp;Elena Zaccheroni,&nbsp;Patricia Hernandez Lopez,&nbsp;Stefania Faletti,&nbsp;Massimiliano Del Bene,&nbsp;Francesco DiMeco,&nbsp;Giuliana Pelicci,&nbsp;Juan M Falcon-Perez","doi":"10.1002/jex2.168","DOIUrl":"10.1002/jex2.168","url":null,"abstract":"<p>Circulating cell-free nucleic acids are considered a promising source of biomarkers for diseases and cancer. Liquid biopsy biomarkers for brain tumours represent a major, still unmet, clinical need. In plasma, nucleic acids can be free or be associated with extracellular vesicles (EVs). Here we report an easy and reproducible method to analyse cell-free nucleic acids in plasma and EVs by conventional flow cytometry easy to translate into the clinics. Nucleic acids associated with the EVs or present in plasma samples are stained by Pyronin Y, which is a fluorescent dye that is preferably binding double-stranded nucleic acids. Fluorescent staining of EVs isolated from cell-conditioned media is suitable for DNA and RNA detection by flow cytometry. The nucleic acids are partially protected from degradation by the EVs’ membrane. Additionally, DNA and RNA can be stained in plasma samples and plasma-derived EVs. Remarkably, analysis of plasma from patients and healthy individuals reveals a difference in their nucleic acid profiles. Taken together, our results indicate that the proposed methodology, which is based on conventional direct flow cytometry, is a promising easy tool for plasma nucleic acid analysis.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"3 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11294885/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141891165","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization of ultracentrifugation-based method to enhance the purity and proteomic profiling depth of plasma-derived extracellular vesicles and particles","authors":"Zurong Wan,&nbsp;Jinghua Gu,&nbsp;Uthra Balaji,&nbsp;Linda Bojmar,&nbsp;Henrik Molina,&nbsp;Søren Heissel,&nbsp;Alexandra E. Pagano,&nbsp;Christopher Peralta,&nbsp;Lee Shaashua,&nbsp;Dorina Ismailgeci,&nbsp;Hope K. Narozniak,&nbsp;Yi Song,&nbsp;William R. Jarnagin,&nbsp;David P. Kelsen,&nbsp;Jaqueline Bromberg,&nbsp;Virginia Pascual,&nbsp;Haiying Zhang","doi":"10.1002/jex2.167","DOIUrl":"10.1002/jex2.167","url":null,"abstract":"<p>Circulating extracellular vesicles and particles (EVPs) are being investigated as potential biomarkers for early cancer detection, prognosis, and disease monitoring. However, the suboptimal purity of EVPs isolated from peripheral blood plasma has posed a challenge of in-depth analysis of the EVP proteome. Here, we compared the effectiveness of different methods for isolating EVPs from healthy donor plasma, including ultracentrifugation (UC)-based protocols, phosphatidylserine-Tim4 interaction-based affinity capture (referred to as “PS”), and several commercial kits. Modified UC methods with an additional UC washing or size exclusion chromatography step substantially improved EVP purity and enabled the detection of additional proteins via proteomic mass spectrometry, including many plasma membrane and cytoplasmic proteins involved in vesicular regulation pathways. This improved performance was reproduced in cancer patient plasma specimens, resulting in the identification of a greater number of differentially expressed EVP proteins, thus expanding the range of potential biomarker candidates. However, PS and other commercial kits did not outperform UC-based methods in improving plasma EVP purity. PS yielded abundant contaminating proteins and a biased enrichment for specific EVP subsets, thus unsuitable for proteomic profiling of plasma EVPs. Therefore, we have optimized UC-based protocols for circulating EVP isolation, which enable further in-depth proteomic analysis for biomarker discovery.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"3 7","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11263976/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141753473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of resazurin phenoxazine dye as a highly sensitive cell viability potency assay for natural killer cell-derived extracellular vesicle-based cancer biotherapeutics","authors":"Frederic St-Denis-Bissonnette,&nbsp;Shirley Qiu,&nbsp;Sarah E. Cummings,&nbsp;Melanie Kirkby,&nbsp;Yohannes Haile,&nbsp;Sarah Wassmer,&nbsp;Gauri Muradia,&nbsp;Jelica Mehic,&nbsp;Andrew Stalker,&nbsp;Amit Shrestha,&nbsp;Michele Ardolino,&nbsp;Seung-Hwan Lee,&nbsp;Dylan Burger,&nbsp;Lisheng Wang,&nbsp;Jessie R. Lavoie","doi":"10.1002/jex2.166","DOIUrl":"10.1002/jex2.166","url":null,"abstract":"<p>Natural killer cell-derived extracellular vesicles (NK-EVs) are candidate biotherapeutics against various cancers. However, standardised potency assays are necessary for a reliable assessment of NK-EVs' cytotoxicity. This study aims to thoroughly evaluate a highly sensitive resazurin phenoxazine-based cell viability potency assay (measurement of the cellular redox metabolism) for quantifying the cytotoxicity of NK-EVs against leukaemia K562 cells (suspension model) and breast cancer MDA-MB-231 cells (adherent model) in vitro. The assay was evaluated based on common analytical parameters setforth by regulatory guidelines, including specificity, selectivity,accuracy, precision, linearity, range and stability. Our results revealed that this resazurin-based cell viability potency assay reliably and reproducibly measured a dose-response of NK-EVs’ cytotoxic activity against both cancer models. The assay showed precision with 5% and 20% variation for intra-run and inter-run variability. The assay signal showed specificity and selectivity of NK-EVs against cancer target cells, as evidenced by the diminished viability of cancer cells following a 5-hour treatment with NK-EVs, without any detectable interference or background. The linearity analysis of target cancer cells revealed strong linearity for densities of 5000 K562 and 1000 MDA-MB-231 cells per test with a consistent range. Importantly, NK-EVs’ dose-response for cytotoxicity showed a strong correlation (|<i>ρ</i>| ∼ 0.8) with the levels of known cytotoxic factors associated with the NK-EVs’ corona (FasL, GNLY, GzmB, PFN and IFN-γ), thereby validating the accuracy of the assay. The assay also distinguished cytotoxicity changes in degraded NK-EVs, indicating the ability of the assay to detect the potential loss of sample integrity. Compared to other commonly reported bioassays (i.e., flow cytometry, cell counting, lactate dehydrogenase release assay, DNA-binding reporter assay and confluence assay), our results support this highly sensitive resazurin-based viability potency assay as a high-throughput and quantitative method for assessing NK-EVs’ cytotoxicity against both suspension and adherent cancer models for evaluating NK-EVs’ biotherapeutics.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"3 7","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11253028/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141636023","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mechanical property estimation of sarcoma-relevant extracellular vesicles using transmission electron microscopy","authors":"Premanshu Kumar Singh,&nbsp;Patricia Sarchet,&nbsp;Catherine Hord,&nbsp;Lucia Casadei,&nbsp;Raphael Pollock,&nbsp;Shaurya Prakash","doi":"10.1002/jex2.158","DOIUrl":"10.1002/jex2.158","url":null,"abstract":"<p>Analysis of single extracellular vesicles (EVs) has the potential to yield valuable label-free information on their morphological structure, biomarkers and therapeutic targets, though such analysis is hindered by the lack of reliable and quantitative measurements of the mechanical properties of these compliant nanoscale particles. The technical challenge in mechanical property measurements arises from the existing tools and methods that offer limited throughput, and the reported elastic moduli range over several orders of magnitude. Here, we report on a flow-based method complemented by transmission electron microscopy (TEM) imaging to provide a high throughput, whole EV deformation analysis for estimating the mechanical properties of liposarcoma-derived EVs as a function of their size. Our study includes extracting morphological data of EVs from a large dataset of 432 TEM images, with images containing single to multiple EVs, and implementing the thin-shell deformation theory. We estimated the elastic modulus, <i>E</i> = 0.16 ± 0.02 MPa (mean±SE) for small EVs (sEVs; 30–150 nm) and <i>E</i> = 0.17 ± 0.03 MPa (mean±SE) for large EVs (<i>l</i>EVs; &gt;150 nm). To our knowledge, this is the first report on the mechanical property estimation of LPS-derived EVs and has the potential to establish a relationship between EV size and EV mechanical properties.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"3 7","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11222873/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141536086","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterisation of extracellular vesicles in baculovirus infection of Spodoptera frugiperda cells","authors":"Lex J. C. Van Es,&nbsp;Robert D. Possee,&nbsp;Linda A. King","doi":"10.1002/jex2.163","DOIUrl":"10.1002/jex2.163","url":null,"abstract":"<p><i>Autographa californica</i> multiple nucleopolyhedrovirus (AcMNPV) is an enveloped DNA virus of the <i>Baculoviridae</i> family. This baculovirus is widely exploited for the biological control of insect pest species and as an expression platform to produce recombinant proteins in insect cells. Extracellular vesicles (EVs) are secreted by all cells and are involved in key roles in many biological processes through their cargo consisting of proteins, RNA or DNA. In viral infections, EVs have been found to transfer both viral and cellular cargo that can elicit either a pro- or antiviral response in recipient cells. Here, small EVs (sEVs) released by <i>Spodoptera frugiperda</i> (Sf) insect cells were characterised for the first time. Using <i>S. frugiperda</i> (SfC1B5) cells stably expressing the baculovirus <i>gp64</i>, the viral envelope protein GP64 was shown to be incorporated into sEVs. Sf9 cells were also transfected with a bacmid AcMNPV genome lacking <i>p6.9</i> (AcΔP6.9) to prevent budded virus production. The protein content of sEVs from both mock- and AcΔP6.9-transfected cells were analysed by mass spectrometry. In addition to GP64, viral proteins Ac-F, ME-53 and viral ubiquitin were identified, as well as many host proteins including TSG101—which may be useful as a protein marker for sEVs.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"3 7","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11212295/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141473245","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}