{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T10:18:29Z","timestamp":1788257909215,"version":"build-2803163510"},"reference-count":66,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T00:00:00Z","timestamp":1788134400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"MUR\u2014DM","award":["118\/2023"],"award-info":[{"award-number":["118\/2023"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Extracellular vesicles (EVs) from Gram-positive bacteria are gaining more interest as potential biotechnological systems for vaccines and drug delivery. However, scalable production protocols remain poorly defined. Here, we evaluated upstream and downstream processes for the controlled production of extracellular vesicles from Staphylococcus epidermidis. Bacterial growth and vesicle release were compared between shake-flask cultures and a controlled stirred-tank bioreactor (STBR), and vesicles were isolated using either ultracentrifugation (UC) or ultrafiltration (UF). STBR cultivation significantly increased biomass accumulation during the exponential phase while preserving the same growth observed in shake flasks. Nanoparticle tracking analysis (NTA) showed time-dependent particle accumulation in both systems, with UF consistently yielding higher particle recovery than UC. Despite differences in yield, vesicles isolated by both methods displayed comparable size distributions and TEM imaging confirmed the spherical morphology. Protein quantification revealed greater protein recovery in UF preparations, whereas particle-to-protein ratios indicated similar sample composition between isolation methods. Together, these results suggest that controlled STBR fermentation combined with UF enables a reproducible, scalable production of S. epidermidis EVs without compromising vesicle integrity. This work provides an integrated framework for Gram-positive EV manufacturing and supports future translational applications.<\/jats:p>","DOI":"10.3390\/ijms27177797","type":"journal-article","created":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T09:48:36Z","timestamp":1788256116000},"page":"7797","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Towards a Scalable Production of Staphylococcus epidermidis Extracellular Vesicles: Integration of Stirred-Tank Bioreactor and Ultrafiltration-Based Recovery"],"prefix":"10.3390","volume":"27","author":[{"given":"Giacomo","family":"Presutti","sequence":"first","affiliation":[{"name":"Department of Applied Science and Technology, Politecnico di Torino, 24 Corso Duca Degli Abruzzi, 10129 Torino, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5249-5426","authenticated-orcid":false,"given":"Francesca","family":"Bosco","sequence":"additional","affiliation":[{"name":"Department of Applied Science and Technology, Politecnico di Torino, 24 Corso Duca Degli Abruzzi, 10129 Torino, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3230-8427","authenticated-orcid":false,"given":"Alessandro","family":"Chiad\u00f2","sequence":"additional","affiliation":[{"name":"Department of Applied Science and Technology, Politecnico di Torino, 24 Corso Duca Degli Abruzzi, 10129 Torino, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5510-5561","authenticated-orcid":false,"given":"Tania","family":"Limongi","sequence":"additional","affiliation":[{"name":"Department of Drug Science and Technology, University of Torino, Via Pietro Giuria 9, 10125 Torino, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1465-3574","authenticated-orcid":false,"given":"Marta","family":"Vallino","sequence":"additional","affiliation":[{"name":"Consiglio Nazionale delle Ricerche (CNR) di Torino, Strada delle Cacce 73, 10129 Torino, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"formal-analysis","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6990-3126","authenticated-orcid":false,"given":"Roberto","family":"Pisano","sequence":"additional","affiliation":[{"name":"Department of Applied Science and Technology, Politecnico di Torino, 24 Corso Duca Degli Abruzzi, 10129 Torino, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.1016\/j.tibtech.2022.03.005","article-title":"The tremendous biomedical potential of bacterial extracellular vesicles","volume":"40","author":"Xie","year":"2022","journal-title":"Trends Biotechnol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"332","DOI":"10.1016\/j.addr.2020.04.004","article-title":"Extracellular vesicles as drug delivery systems: Why and how?","volume":"159","author":"Elsharkasy","year":"2020","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"620","DOI":"10.1038\/nrmicro3480","article-title":"Through the wall: Extracellular vesicles in Gram-positive bacteria, mycobacteria and fungi","volume":"13","author":"Brown","year":"2015","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Liu, Y., Defourny, K.A.Y., Smid, E.J., and Abee, T. (2018). Gram-positive bacterial extracellular vesicles and their impact on health and disease. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.01502"},{"key":"ref_5","first-page":"169","article-title":"Bacterial extracellular vesicles as bioactive nanocarriers for drug delivery: Advances and perspectives","volume":"14","author":"Liu","year":"2022","journal-title":"Bioact. Mater."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Pezzotti, G., Adachi, T., Imamura, H., Bristol, D.R., Adachi, K., Yamamoto, T., Kanamura, N., Marin, E., Zhu, W., and Kawai, T. (2023). In Situ Raman Study of Neurodegenerated Human Neuroblastoma Cells Exposed to Outer-Membrane Vesicles Isolated from Porphyromonas gingivalis. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241713351"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"5425","DOI":"10.1002\/pmic.200900338","article-title":"Gram-positive bacteria produce membrane vesicles: Proteomics-based characterization of Staphylococcus aureus-derived membrane vesicles","volume":"9","author":"Lee","year":"2009","journal-title":"Proteomics"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1795","DOI":"10.1007\/s00253-021-11140-1","article-title":"Characterization and function of membrane vesicles in Gram-positive bacteria","volume":"105","author":"Cao","year":"2021","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"127612","DOI":"10.1016\/j.micres.2024.127612","article-title":"Extracellular vesicles from Staphylococcus aureus promote the pathogenicity of Pseudomonas aeruginosa","volume":"281","author":"Subsomwong","year":"2024","journal-title":"Microbiol. Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1379","DOI":"10.1038\/s41467-018-03847-z","article-title":"Release of Staphylococcus aureus extracellular vesicles and their application as a vaccine platform","volume":"9","author":"Wang","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1","DOI":"10.20517\/evcna.2024.76","article-title":"Consistency in bacterial extracellular vesicle production: Key to their application in human health","volume":"6","author":"Dai","year":"2025","journal-title":"Extracell. Vesicles Circ. Nucl. Acids"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1038\/s41579-018-0112-2","article-title":"Types and origins of bacterial membrane vesicles","volume":"17","author":"Toyofuku","year":"2019","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"106167","DOI":"10.1016\/j.micpath.2023.106167","article-title":"Correlation between bacterial extracellular vesicles and antibiotics: A potentially antibacterial strategy","volume":"181","author":"Yang","year":"2023","journal-title":"Microb. Pathog."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e00433-20","DOI":"10.1128\/IAI.00433-20","article-title":"Extracellular vesicle biogenesis and functions in gram-positive bacteria","volume":"88","author":"Briaud","year":"2020","journal-title":"Infect. Immun."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e84","DOI":"10.1002\/jex2.84","article-title":"A new isolation method for bacterial extracellular vesicles providing greater purity and improved proteomic detection of vesicle proteins","volume":"2","author":"Le","year":"2023","journal-title":"J. Extracell. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e00676-21","DOI":"10.1128\/mSphere.00676-21","article-title":"Temperature Influences the Composition and Cytotoxicity of Extracellular Vesicles in Staphylococcus aureus","volume":"6","author":"Briaud","year":"2021","journal-title":"mSphere"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e70077","DOI":"10.1002\/jev2.70077","article-title":"Beta-Lactam Antibiotics Promote Extracellular Vesicle Production of Staphylococcus aureus Through ROS-Mediated Lipid Metabolic Reprogramming","volume":"14","author":"Huang","year":"2025","journal-title":"J. Extracell. Vesicles"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Torabian, P., Singh, N., Crawford, J., Gonzalez, G., Burgado, N., Videva, M., Miller, A., Perdue, J., Dinu, M., and Pietropaoli, A. (2023). The effect of clinically relevant beta-lactam, aminoglycoside, and quinolone antibiotics on bacterial extracellular vesicle release from E. coli. bioRxiv.","DOI":"10.1101\/2023.11.22.568081"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Zaborowska, M., Taul\u00e9 Flores, C., Vazirisani, F., Shah, F.A., Thomsen, P., and Trobos, M. (2020). Extracellular Vesicles Influence the Growth and Adhesion of Staphylococcus epidermidis Under Antimicrobial Selective Pressure. Front. Microbiol., 11.","DOI":"10.3389\/fmicb.2020.01132"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e70001","DOI":"10.1002\/elsc.70001","article-title":"Pitfalls in Early Bioprocess Development Using Shake Flask Cultivations","volume":"25","author":"Brauneck","year":"2025","journal-title":"Eng. Life Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/j.biotechadv.2008.10.006","article-title":"Bioreactor scale-up and oxygen transfer rate in microbial processes: An overview","volume":"27","author":"Gomez","year":"2009","journal-title":"Biotechnol. Adv."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"6978","DOI":"10.1016\/j.vaccine.2019.01.076","article-title":"Spontaneously released Neisseria meningitidis outer membrane vesicles as vaccine platform: Production and purification","volume":"37","author":"Gerritzen","year":"2019","journal-title":"Vaccine"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"101978","DOI":"10.1016\/j.scr.2020.101978","article-title":"Strategies for scalable manufacturing and translation of MSC-derived extracellular vesicles","volume":"48","author":"Adlerz","year":"2020","journal-title":"Stem Cell Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1016\/j.molmed.2018.01.006","article-title":"Manufacturing Exosomes: A Promising Therapeutic Platform","volume":"24","author":"Colao","year":"2018","journal-title":"Trends Mol. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e2506011","DOI":"10.4014\/jmb.2506.06011","article-title":"Standardizing Bacterial Extracellular Vesicle Purification: A Call for Consensus","volume":"35","author":"Choi","year":"2025","journal-title":"J. Microbiol. Biotechnol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1002\/elps.202200133","article-title":"Production and purification of bacterial membrane vesicles for biotechnology applications: Challenges and opportunities","volume":"44","author":"Santacruz","year":"2023","journal-title":"Electrophoresis"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"461773","DOI":"10.1016\/j.chroma.2020.461773","article-title":"Modern isolation and separation techniques for extracellular vesicles","volume":"1636","author":"Liangsupree","year":"2021","journal-title":"J. Chromatogr. A"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1800528","DOI":"10.1002\/biot.201800528","article-title":"Scalable Production and Isolation of Extracellular Vesicles: Available Sources and Lessons from Current Industrial Bioprocesses","volume":"14","author":"Paganini","year":"2019","journal-title":"Biotechnol. J."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1066","DOI":"10.1016\/j.tibtech.2020.05.012","article-title":"Technologies and Standardization in Research on Extracellular Vesicles","volume":"38","author":"Gandham","year":"2020","journal-title":"Trends Biotechnol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1007\/s41048-019-00095-6","article-title":"Natural and engineered bacterial outer membrane vesicles","volume":"5","author":"Qing","year":"2019","journal-title":"Biophys. Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"29509","DOI":"10.3402\/jev.v4.29509","article-title":"High-speed centrifugation induces aggregation of extracellular vesicles","volume":"4","author":"Linares","year":"2015","journal-title":"J. Extracell. Vesicles"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3036","DOI":"10.1002\/elps.201800526","article-title":"Recent advances and challenges in the recovery and purification of cellular exosomes","volume":"40","year":"2019","journal-title":"Electrophoresis"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1002\/prca.200800093","article-title":"Microfiltration isolation of human urinary exosomes for characterization by MS","volume":"4","author":"Merchant","year":"2010","journal-title":"Proteom. Clin. Appl."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"e70362","DOI":"10.1002\/wer.70362","article-title":"Membrane Fouling in Vibratory MBRs: Impact Distance Complements Wall Shear Stress as a Fouling Predictor","volume":"98","author":"Kaya","year":"2026","journal-title":"Water Environ. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"17261","DOI":"10.1021\/acs.analchem.5c03781","article-title":"Optimized Tangential Flow Filtration for Efficient Isolation of Highly Pure and Bioactive sEVs","volume":"97","author":"Luo","year":"2025","journal-title":"Anal. Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1016\/j.ijpharm.2017.03.046","article-title":"Analytical ultracentrifugation for analysis of doxorubicin loaded liposomes","volume":"523","author":"Mehn","year":"2017","journal-title":"Int. J. Pharm."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"S82","DOI":"10.1016\/S0263-8762(97)80006-8","article-title":"Sedimentation and fluidisation: Part I","volume":"75","author":"Richardson","year":"1997","journal-title":"Chem. Eng. Res. Des."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2401","DOI":"10.1016\/j.jcp.2009.11.041","article-title":"Simulation of concentrated suspensions using the force-coupling method","volume":"229","author":"Yeo","year":"2010","journal-title":"J. Comput. Phys."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"218301","DOI":"10.1103\/PhysRevLett.111.218301","article-title":"Discontinuous shear thickening of frictional hard-sphere suspensions","volume":"111","author":"Seto","year":"2013","journal-title":"Phys. Rev. Lett."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Busatto, S., Vilanilam, G., Ticer, T., Lin, W.L., Dickson, D.W., Shapiro, S., Bergese, P., and Wolfram, J. (2018). Tangential flow filtration for highly efficient concentration of extracellular vesicles from large volumes of fluid. Cells, 7.","DOI":"10.3390\/cells7120273"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Torres-Vanegas, J.D., Rincon-Tellez, N., Guzm\u00e1n-Sastoque, P., Valderrama-Rincon, J.D., Cruz, J.C., and Reyes, L.H. (2024). Production and purification of outer membrane vesicles encapsulating green fluorescent protein from Escherichia coli: A step towards scalable OMV technologies. Front. Bioeng. Biotechnol., 12.","DOI":"10.3389\/fbioe.2024.1436352"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e00084","DOI":"10.1002\/adtp.202500084","article-title":"Optimization of the Isolation Method for Large-Scale Production of Synthetic Bacterial Vesicles for Cancer Immunotherapy","volume":"8","author":"Ordouzadeh","year":"2025","journal-title":"Adv. Ther."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"9401","DOI":"10.1007\/s00253-019-10163-z","article-title":"Continuous production of Neisseria meningitidis outer membrane vesicles","volume":"103","author":"Gerritzen","year":"2019","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"G\u00f3mez-Ch\u00e1vez, F., Cedillo-Pel\u00e1ez, C., Zapi-Col\u00edn, L.A., Guti\u00e9rrez-Gonz\u00e1lez, G., Mart\u00ednez-Torres, I., Peralta, H., Chavez-Galan, L., Avila-Calder\u00f3n, E.D., Contreras-Rodr\u00edguez, A., and Bartolo-Aguilar, Y. (2021). The extracellular vesicles from the commensal Staphylococcus epidermidis atcc12228 strain regulate skin inflammation in the imiquimod-induced psoriasis murine model. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222313029"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1016\/j.jid.2023.02.023","article-title":"Extracellular Vesicles of Commensal Skin Microbiota Alleviate Cutaneous Inflammation in Atopic Dermatitis Mouse Model by Re-Establishing Skin Homeostasis","volume":"145","author":"Zhou","year":"2025","journal-title":"J. Investig. Dermatol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"126067","DOI":"10.1016\/j.ijpharm.2025.126067","article-title":"The inhibitory effect of extracellular vesicles derived from S. epidermidis on MRSA biofilms","volume":"683","author":"Xian","year":"2025","journal-title":"Int. J. Pharm."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"2197","DOI":"10.1016\/j.apsb.2025.02.038","article-title":"Microneedle delivery platform integrated with Staphylococcus epidermidis-derived extracellular vesicles-based nanoantibiotics for efficient bacterial infection atopic dermatitis treatment","volume":"15","author":"Zhou","year":"2025","journal-title":"Acta Pharm. Sin. B"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"2300269","DOI":"10.1002\/pmic.202300269","article-title":"The effect of altered pH growth conditions on the production, composition, and proteomes of Helicobacter pylori outer membrane vesicles","volume":"24","author":"Johnston","year":"2024","journal-title":"Proteomics"},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Yuan, Z., Huang, W., Chan, P., Zhou, J., Liang, J., and Guo, L. (2025). Membrane Vesicles of Enterococcus faecalis: In Vitro Composition Analysis and Macrophage Inflammatory Response Under Different pH Conditions. Microorganisms, 13.","DOI":"10.3390\/microorganisms13061344"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Iwabuchi, Y., Nakamura, T., Kusumoto, Y., Nakao, R., Iwamoto, T., Shinozuka, O., and Senpuku, H. (2021). Effects of pH on the properties of membrane vesicles including glucosyltransferase in Streptococcus mutans. Microorganisms, 9.","DOI":"10.3390\/microorganisms9112308"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"19861","DOI":"10.3402\/jev.v2i0.19861","article-title":"How pure are your vesicles?","volume":"2","author":"Webber","year":"2013","journal-title":"J. Extracell. Vesicles"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"e00697-18","DOI":"10.1128\/JB.00697-18","article-title":"Lag Phase Is a Dynamic, Organized, Adaptive, and Evolvable Period That Prepares Bacteria for Cell Division","volume":"201","author":"Bertrand","year":"2019","journal-title":"J. Bacteriol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"e12002","DOI":"10.1002\/jev2.12002","article-title":"Hypoxia and extracellular vesicles: A review on methods, vesicular cargo and functions","volume":"10","author":"Bister","year":"2020","journal-title":"J. Extracell. Vesicles"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1007\/s10616-015-9913-6","article-title":"Systematic review of factors influencing extracellular vesicle yield from cell cultures","volume":"68","author":"Gudbergsson","year":"2016","journal-title":"Cytotechnology"},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Jaishankar, J., and Srivastava, P. (2017). Molecular basis of stationary phase survival and applications. Front. Microbiol., 8.","DOI":"10.3389\/fmicb.2017.02000"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1186\/s13287-025-04341-2","article-title":"Continuous collection of human mesenchymal-stromal-cell-derived extracellular vesicles from a stirred tank reactor operated under xenogeneic-free conditions for therapeutic applications","volume":"16","author":"Ulpiano","year":"2025","journal-title":"Stem Cell Res. Ther."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"2561","DOI":"10.1016\/j.vaccine.2021.03.059","article-title":"Vaccination with meningococcal outer membrane vesicles carrying Borrelia OspA protects against experimental Lyme borreliosis","volume":"39","author":"Klouwens","year":"2021","journal-title":"Vaccine"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"879","DOI":"10.1016\/j.nano.2015.01.003","article-title":"Ultrafiltration with size-exclusion liquid chromatography for high yield isolation of extracellular vesicles preserving intact biophysical and functional properties","volume":"11","author":"Nordin","year":"2015","journal-title":"Nanomedicine"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"27031","DOI":"10.3402\/jev.v4.27031","article-title":"Optimized exosome isolation protocol for cell culture supernatant and human plasma","volume":"4","author":"Lobb","year":"2015","journal-title":"J. Extracell. Vesicles"},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Cannizzaro, F., Gallo, A., La Scala, S., Gallo, G., and Faddetta, T. (2026). Bacterial Extracellular Vesicles in Biotechnology: Current Challenges and Strategies for Production Enhancement. Fermentation, 12.","DOI":"10.3390\/fermentation12020086"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"678","DOI":"10.1186\/s12951-025-03768-2","article-title":"Advances of extracellular vesicles isolation and detection frontier technology: From heterogeneity analysis to clinical application","volume":"23","author":"Li","year":"2025","journal-title":"J. Nanobiotechnology"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"1657","DOI":"10.1152\/ajprenal.00434.2006","article-title":"Rapid isolation of urinary exosomal biomarkers using a nanomembrane ultrafiltration concentrator","volume":"292","author":"Cheruvanky","year":"2007","journal-title":"Am. J. Physiol. Ren. Physiol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"121158","DOI":"10.1016\/j.biomaterials.2021.121158","article-title":"Immunomodulatory effects exerted by extracellular vesicles from Staphylococcus epidermidis and Staphylococcus aureus isolated from bone-anchored prostheses","volume":"278","author":"Zaborowska","year":"2021","journal-title":"Biomaterials"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"8545347","DOI":"10.1155\/2018\/8545347","article-title":"Isolation of Extracellular Vesicles: General Methodologies and Latest Trends","volume":"2018","author":"Konoshenko","year":"2018","journal-title":"BioMed Res. Int."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"e12266","DOI":"10.1002\/jev2.12266","article-title":"Comparative analysis of tangential flow filtration and ultracentrifugation, both combined with subsequent size exclusion chromatography, for the isolation of small extracellular vesicles","volume":"11","author":"Visan","year":"2022","journal-title":"J. Extracell. Vesicles"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"5871","DOI":"10.1021\/acsbiomaterials.3c00678","article-title":"Comparative Studies of Different Preservation Methods and Relative Freeze-Drying Formulations for Extracellular Vesicle Pharmaceutical Applications","volume":"9","author":"Susa","year":"2023","journal-title":"ACS Biomater. Sci. Eng."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/17\/7797\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T09:57:36Z","timestamp":1788256656000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/17\/7797"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,31]]},"references-count":66,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["ijms27177797"],"URL":"https:\/\/doi.org\/10.3390\/ijms27177797","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,31]]}}}