{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T19:51:55Z","timestamp":1770493915188,"version":"3.49.0"},"reference-count":83,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2023,3,15]],"date-time":"2023-03-15T00:00:00Z","timestamp":1678838400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","award":["PTDC\/BTM-ORG\/32187\/2017"],"award-info":[{"award-number":["PTDC\/BTM-ORG\/32187\/2017"]}]},{"name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","award":["2022.10467.PTDC"],"award-info":[{"award-number":["2022.10467.PTDC"]}]},{"name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","award":["UIDB\/04462\/2020"],"award-info":[{"award-number":["UIDB\/04462\/2020"]}]},{"name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","award":["UIDP\/04462\/2020"],"award-info":[{"award-number":["UIDP\/04462\/2020"]}]},{"name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","award":["LA\/P\/0087\/2020"],"award-info":[{"award-number":["LA\/P\/0087\/2020"]}]},{"name":"FCT\/Minist\u00e9rio da Ci\u00eancia, Tecnologia e Ensino Superior","award":["PTDC\/BTM-ORG\/32187\/2017"],"award-info":[{"award-number":["PTDC\/BTM-ORG\/32187\/2017"]}]},{"name":"FCT\/Minist\u00e9rio da Ci\u00eancia, Tecnologia e Ensino Superior","award":["2022.10467.PTDC"],"award-info":[{"award-number":["2022.10467.PTDC"]}]},{"name":"FCT\/Minist\u00e9rio da Ci\u00eancia, Tecnologia e Ensino Superior","award":["UIDB\/04462\/2020"],"award-info":[{"award-number":["UIDB\/04462\/2020"]}]},{"name":"FCT\/Minist\u00e9rio da Ci\u00eancia, Tecnologia e Ensino Superior","award":["UIDP\/04462\/2020"],"award-info":[{"award-number":["UIDP\/04462\/2020"]}]},{"name":"FCT\/Minist\u00e9rio da Ci\u00eancia, Tecnologia e Ensino Superior","award":["LA\/P\/0087\/2020"],"award-info":[{"award-number":["LA\/P\/0087\/2020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>The short shelf life of platelet concentrates (PC) of up to 5\u20137 days leads to higher wastage due to expiry. To address this massive financial burden on the healthcare system, alternative applications for expired PC have emerged in recent years. Engineered nanocarriers functionalized with platelet membranes have shown excellent targeting abilities for tumor cells owing to their platelet membrane proteins. Nevertheless, synthetic drug delivery strategies have significant drawbacks that platelet-derived extracellular vesicles (pEV) can overcome. We investigated, for the first time, the use of pEV as a carrier of the anti-breast cancer drug paclitaxel, considering it as an appealing alternative to improve the therapeutic potential of expired PC. The pEV released during PC storage showed a typical EV size distribution profile (100\u2013300 nm) with a cup-shaped morphology. Paclitaxel-loaded pEV showed significant anti-cancer effects in vitro, as demonstrated by their anti-migratory (&gt;30%), anti-angiogenic (&gt;30%), and anti-invasive (&gt;70%) properties in distinct cells found in the breast tumor microenvironment. We provide evidence for a novel application for expired PC by suggesting that the field of tumor treatment research may be broadened by the use of natural carriers.<\/jats:p>","DOI":"10.3390\/pharmaceutics15030953","type":"journal-article","created":{"date-parts":[[2023,3,15]],"date-time":"2023-03-15T06:36:36Z","timestamp":1678862196000},"page":"953","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Clinically Expired Platelet Concentrates as a Source of Extracellular Vesicles for Targeted Anti-Cancer Drug Delivery"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8291-6404","authenticated-orcid":false,"given":"Ana","family":"Meliciano","sequence":"first","affiliation":[{"name":"iBET, Instituto de Biologia Experimental e Tecnol\u00f3gica, Apartado 12, 2781-901 Oeiras, Portugal"},{"name":"ITQB-NOVA, Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa, Avenida da Rep\u00fablica, 2780-157 Oeiras, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4033-9020","authenticated-orcid":false,"given":"Daniela","family":"Salvador","sequence":"additional","affiliation":[{"name":"iBET, Instituto de Biologia Experimental e Tecnol\u00f3gica, Apartado 12, 2781-901 Oeiras, Portugal"},{"name":"ITQB-NOVA, Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa, Avenida da Rep\u00fablica, 2780-157 Oeiras, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3850-2909","authenticated-orcid":false,"given":"Pedro","family":"Mendon\u00e7a","sequence":"additional","affiliation":[{"name":"IPST, Instituto Portugu\u00eas do Sangue e da Transplanta\u00e7\u00e3o, 1740-005 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3856-873X","authenticated-orcid":false,"given":"Ana Filipa","family":"Louro","sequence":"additional","affiliation":[{"name":"iBET, Instituto de Biologia Experimental e Tecnol\u00f3gica, Apartado 12, 2781-901 Oeiras, Portugal"},{"name":"ITQB-NOVA, Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa, Avenida da Rep\u00fablica, 2780-157 Oeiras, Portugal"}]},{"given":"Margarida","family":"Serra","sequence":"additional","affiliation":[{"name":"iBET, Instituto de Biologia Experimental e Tecnol\u00f3gica, Apartado 12, 2781-901 Oeiras, Portugal"},{"name":"ITQB-NOVA, Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa, Avenida da Rep\u00fablica, 2780-157 Oeiras, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2023,3,15]]},"reference":[{"key":"ref_1","unstructured":"(2022, November 02). World Health Organization. Available online: https:\/\/apps.who.int\/iris\/handle\/10665\/273826."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"998","DOI":"10.1111\/vox.13095","article-title":"Activity-based cost of platelet transfusions in medical and surgical inpatients at a US hospital","volume":"116","author":"Hofmann","year":"2021","journal-title":"Vox Sang."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1111\/tme.12155","article-title":"Why has demand for platelet components increased? A review","volume":"24","author":"Estcourt","year":"2014","journal-title":"Transfus. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.transci.2014.08.006","article-title":"Multifaceted regenerative lives of expired platelets in the second decade of the 21st century","volume":"51","author":"Burnouf","year":"2014","journal-title":"Transfus. Apher. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1007\/s12663-011-0182-4","article-title":"Platelet concentrates: Past, present and future","volume":"10","author":"Prakash","year":"2011","journal-title":"J. Maxillofac. Oral. Surg."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Salvador, D., Almeida, H., Rego, D., Mendon\u00e7a, P., Sousa, A.P., Serra, M., and Redondo, L. (2022). Pulsed Electric Fields for Valorization of Platelets with No Therapeutic Value towards a High Biomedical Potential Product\u2014A Proof of Concept. Appl. Sci., 12.","DOI":"10.3390\/app12125773"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1186\/s13287-018-0863-8","article-title":"Human platelet lysate in mesenchymal stromal cell expansion according to a GMP grade protocol: A cell factory experience","volume":"9","author":"Becherucci","year":"2018","journal-title":"Stem. Cell Res. Ther."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2355","DOI":"10.17576\/jsm-2021-5008-18","article-title":"Expired platelet concentrate as a source of human platelet lysate for xenogeneic-free culture of human dermal fibroblasts","volume":"50","author":"Hassan","year":"2021","journal-title":"Sains Malays."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1080\/09537104.2018.1445840","article-title":"Comparison of human platelet lysate alternatives using expired and freshly isolated platelet concentrates for adipose-derived stromal cell expansion","volume":"30","author":"Dessels","year":"2019","journal-title":"Platelets"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"598","DOI":"10.1016\/j.tibtech.2020.10.004","article-title":"Prospective Therapeutic Applications of Platelet Extracellular Vesicles","volume":"39","author":"Johnson","year":"2021","journal-title":"Trends Biotechnol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Woith, E., Fuhrmann, G., and Melzig, M.F. (2019). Extracellular Vesicles-Connecting Kingdoms. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20225695"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s11060-013-1084-8","article-title":"Biogenesis of extracellular vesicles (EV): Exosomes, microvesicles, retrovirus-like vesicles, and apoptotic bodies","volume":"113","author":"Akers","year":"2013","journal-title":"J. Neurooncol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1007\/s11010-020-03703-z","article-title":"Inside(sight) of tiny communicator: Exosome biogenesis, secretion, and uptake","volume":"467","author":"Jadli","year":"2020","journal-title":"Mol. Cell Biochem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1038\/s41556-018-0049-8","article-title":"Size matters in nanoscale communication","volume":"20","author":"Zijlstra","year":"2018","journal-title":"Nat. Cell Biol."},{"key":"ref_15","first-page":"206","article-title":"The release of vesicles from platelets following adhesion to vessel walls in vitro","volume":"53","author":"Warren","year":"1972","journal-title":"Br. J. Exp. Pathol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1182\/blood.V77.4.887.887","article-title":"Vesiculation of platelets during in vitro aging","volume":"77","author":"Bode","year":"1991","journal-title":"Blood"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"578","DOI":"10.1097\/MOH.0b013e32833e77ee","article-title":"Clinical relevance of microparticles from platelets and megakaryocytes","volume":"17","author":"Italiano","year":"2010","journal-title":"Curr. Opin. Hematol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1038\/s41565-021-00931-2","article-title":"Extracellular vesicles as a next-generation drug delivery platform","volume":"16","author":"Herrmann","year":"2021","journal-title":"Nat. Nanotechnol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"9","DOI":"10.4137\/CGM.S11285","article-title":"Regulation of tumor growth and metastasis: The role of tumor microenvironment","volume":"7","author":"Goubran","year":"2014","journal-title":"Cancer Growth Metastasis"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1080\/10717544.2020.1748758","article-title":"Prospects and challenges of extracellular vesicle-based drug delivery system: Considering cell source","volume":"27","author":"Meng","year":"2020","journal-title":"Drug Deliv."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.biomaterials.2015.10.046","article-title":"Targeted drug delivery to circulating tumor cells via platelet membrane-functionalized particles","volume":"76","author":"Li","year":"2016","journal-title":"Biomaterials"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"10151","DOI":"10.2147\/IJN.S285952","article-title":"Platelet-Mimicking Drug Delivery Nanoparticles for Enhanced Chemo-Photothermal Therapy of Breast Cancer","volume":"15","author":"Pei","year":"2020","journal-title":"Int. J. Nanomed."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2683","DOI":"10.7150\/thno.23654","article-title":"Platelet-camouflaged nanococktail: Simultaneous inhibition of drug-resistant tumor growth and metastasis via a cancer cells and tumor vasculature dual-targeting strategy","volume":"8","author":"Jing","year":"2018","journal-title":"Theranostics"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Bang, K.H., Na, Y.G., Huh, H.W., Hwang, S.J., Kim, M.S., Kim, M., Lee, H.K., and Cho, C.W. (2019). The Delivery Strategy of Paclitaxel Nanostructured Lipid Carrier Coated with Platelet Membrane. Cancers, 11.","DOI":"10.3390\/cancers11060807"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"46218","DOI":"10.1039\/C5RA05339K","article-title":"Targeted killing of metastatic cells using a platelet-inspired drug delivery system","volume":"5","author":"Pan","year":"2015","journal-title":"RSC Adv."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1186\/s12929-020-00633-2","article-title":"Clinical-grade cryopreserved doxorubicin-loaded platelets: Role of cancer cells and platelet extracellular vesicles activation loop","volume":"27","author":"Wu","year":"2020","journal-title":"J. Biomed. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1004","DOI":"10.1056\/NEJM199504133321507","article-title":"Paclitaxel (taxol)","volume":"11","author":"Rowinsky","year":"1995","journal-title":"N. Engl. J. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"626910","DOI":"10.3389\/fcell.2021.626910","article-title":"Targeting Strategies for Enhancing Paclitaxel Specificity in Chemotherapy","volume":"9","author":"Ma","year":"2021","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_29","unstructured":"EDQM, 20th Edition of the Guide to the Preparation, Use and Quality Assurance of Blood Components (2022, November 08). European Committee on Blood Transfusion (CD-P-TS). Available online: https:\/\/www.edqm.eu\/en\/blood-guide."},{"key":"ref_30","unstructured":"(2022, November 08). Dire\u00e7\u00e3o-Geral da Sa\u00fade, Utiliza\u00e7\u00e3o Cl\u00ednica de Concentrados Plaquet\u00e1rios no Adulto M\u00e9dicos do Sistema Nacional de Sa\u00fade. Available online: www.dgs.pt."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1006\/meth.2000.1006","article-title":"Evaluation of platelet function by flow cytometry","volume":"21","author":"Michelson","year":"2000","journal-title":"Methods"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1007\/s10753-018-0893-5","article-title":"Ability of Platelet-Derived Extracellular Vesicles to Promote Neutrophil-Endothelial Cell Interactions","volume":"42","author":"Kuravi","year":"2019","journal-title":"Inflammation"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2104296","DOI":"10.1002\/advs.202104296","article-title":"Bioactivity and miRNome Profiling of Native Extracellular Vesicles in Human Induced Pluripotent Stem Cell-Cardiomyocyte Differentiation","volume":"9","author":"Louro","year":"2022","journal-title":"Adv. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"24858","DOI":"10.3402\/jev.v3.24858","article-title":"The impact of disparate isolation methods for extracellular vesicles on downstream RNA profiling","volume":"3","author":"Deun","year":"2014","journal-title":"J. Extracell. Vesicles"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"655","DOI":"10.1016\/j.nano.2015.10.012","article-title":"Development of exosome-encapsulated paclitaxel to overcome MDR in cancer cells","volume":"12","author":"Kim","year":"2016","journal-title":"Nanomedicine"},{"key":"ref_36","first-page":"29","article-title":"Spectrophotometric estimation of paclitaxel","volume":"2","author":"Kesarwani","year":"2011","journal-title":"Int. J. Adv. Pharm. Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e3","DOI":"10.7717\/peerj-achem.3","article-title":"A simple spectrophotometric evaluation method for the hydrophobic anticancer drug paclitaxel","volume":"2","author":"Sugo","year":"2020","journal-title":"PeerJ Anal. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"113390","DOI":"10.1016\/j.ab.2019.113390","article-title":"Comparison of methods to isolate proteins from extracellular vesicles for mass spectrometry-based proteomic analyses","volume":"584","author":"Subedi","year":"2019","journal-title":"Anal. Biochem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1311","DOI":"10.1038\/nprot.2012.065","article-title":"Fluorescent labeling of nano-sized vesicles released by cells and subsequent quantitative and qualitative analysis by high-resolution flow cytometry","volume":"7","author":"Stoorvogel","year":"2012","journal-title":"Nat. Protoc."},{"key":"ref_40","unstructured":"Carpentier, G., and ImageJ contribution: Angiogenesis Analyzer (2021, January 11). ImageJ News. Available online: http:\/\/image.bio.methods.free.fr\/ImageJ\/?Angiogenesis-Analyzer-for-ImageJ&lang=en."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Suarez-Arnedo, A., Torres Figueroa, F., Clavijo, C., Arbel\u00e1ez, P., Cruz, J.C., and Mu\u00f1oz-Camargo, C. (2020). An image J plugin for the high throughput image analysis of in vitro scratch wound healing assays. PLoS ONE, 15.","DOI":"10.1101\/2020.04.20.050831"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1295","DOI":"10.1021\/acs.bioconjchem.2c00197","article-title":"DNA Origami-Platelet Adducts: Nanoconstruct 993 Binding without Platelet Activation","volume":"33","author":"Walawalkar","year":"2022","journal-title":"Bioconjug. Chem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1038\/s41598-020-57497-7","article-title":"A comparison of methods for the isolation and separation of extracellular vesicles from protein and lipid particles in human serum","volume":"10","author":"Brennan","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2873","DOI":"10.1007\/s00018-018-2773-4","article-title":"Detailed analysis of the plasma extracellular vesicle proteome after separation from lipoproteins","volume":"75","author":"Karimi","year":"2018","journal-title":"Cell Mol. Life Sci."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"32945","DOI":"10.3402\/jev.v5.32945","article-title":"Techniques used for the isolation and characterization of extracellular vesicles: Results of a worldwide survey","volume":"5","author":"Gardiner","year":"2016","journal-title":"J. Extracell. Vesicles"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"712","DOI":"10.1016\/j.nano.2011.09.006","article-title":"Quantitative and qualitative flow cytometric analysis of nanosized cell-derived membrane vesicles","volume":"8","author":"Aalberts","year":"2012","journal-title":"Nanomedicine"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1016\/0167-7799(94)90120-1","article-title":"Taxol: The chemistry and structure-activity relationships of a novel anticancer agent","volume":"12","author":"Kingston","year":"1994","journal-title":"Trends Biotechnol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1116","DOI":"10.3389\/fphar.2018.01116","article-title":"A New Approach for Loading Anticancer Drugs Into Mesenchymal Stem Cell-Derived Exosome Mimetics for Cancer Therapy","volume":"9","author":"Kalimuthu","year":"2018","journal-title":"Front. Pharmacol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1519","DOI":"10.2217\/nnm-2021-0123","article-title":"Extracellular vesicles as drug vectors for precise cancer treatment","volume":"16","author":"Cao","year":"2021","journal-title":"Nanomedicine"},{"key":"ref_50","first-page":"1843","article-title":"The microtubule-affecting drug paclitaxel has antiangiogenic activity","volume":"2","author":"Belotti","year":"1996","journal-title":"Clin. Cancer Res."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1007\/s10456-013-9334-0","article-title":"The pharmacological bases of the antiangiogenic activity of paclitaxel","volume":"16","author":"Bocci","year":"2013","journal-title":"Angiogenesis"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"2799","DOI":"10.7150\/ijms.43101","article-title":"A high expression ratio of RhoA\/RhoB is associated with the migratory and invasive properties of basal-like Breast Tumors","volume":"17","author":"Privat","year":"2020","journal-title":"Int. J. Med. Sci."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"68803","DOI":"10.18632\/oncotarget.11931","article-title":"A highly invasive subpopulation of MDA-MB-231 breast cancer cells shows accelerated growth, differential chemoresistance, features of apocrine tumors and reduced tumorigenicity in vivo","volume":"7","author":"Amaro","year":"2016","journal-title":"Oncotarget"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/j.transci.2009.07.005","article-title":"Platelet utilization in the developing world: Strategies to optimize platelet transfusion practices","volume":"41","author":"Verma","year":"2009","journal-title":"Transfus. Apher. Sci."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1120","DOI":"10.1016\/0140-6736(92)93150-L","article-title":"Angiogenesis, assessed by platelet\/endothelial cell adhesion molecule antibodies, as indicator of node metastases and survival in breast cancer","volume":"340","author":"Horak","year":"1992","journal-title":"Lancet"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1134","DOI":"10.1016\/S0140-6736(16)31891-8","article-title":"Breast cancer","volume":"389","author":"Harbeck","year":"2017","journal-title":"Lancet"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"2395","DOI":"10.1111\/trf.14883","article-title":"Geration of procoagulant collagen- and thrombin- activated platelets in platelet concentrates derived from buffy coat: The role of processing, pathogen inactivation, and storage","volume":"58","author":"Calderara","year":"2018","journal-title":"Transfusion"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1026386","DOI":"10.3389\/fphar.2022.1026386","article-title":"Platelets and platelet extracellular vesicles in drug delivery therapy: A review of the current status and future prospects","volume":"13","author":"Dai","year":"2022","journal-title":"Front. Pharmacol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"1535750","DOI":"10.1080\/20013078.2018.1535750","article-title":"Minimal information for studies of extracellular vesicles 2018 (MISEV2018): A position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines","volume":"7","author":"Witwer","year":"2018","journal-title":"J. Extracell. Vesicles"},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Jung, M.K., and Mun, J.Y. (2018). Sample Preparation and Imaging of exosomes by Transmission Electron Microscopy. J. Vis. Exp., e56482.","DOI":"10.3791\/56482-v"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"24692","DOI":"10.3402\/jev.v3.24692","article-title":"Isolation and characterization of platelet-derived extracellular vesicles","volume":"3","author":"Aatonen","year":"2014","journal-title":"J. Extracell. Vesicles"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"19159","DOI":"10.1038\/s41598-019-55477-0","article-title":"Optimized Isolation of Extracellular Vesicles From Various Organic Sources Using Aqueous Two-Phase System","volume":"9","author":"Bozkurt","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"e2000118","DOI":"10.1002\/pmic.202000118","article-title":"Understanding extracellular vesicle and nanoparticle heterogeneity: Novel methods and considerations","volume":"21","author":"Phillips","year":"2021","journal-title":"Proteomics"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"1479","DOI":"10.3389\/fphys.2018.01479","article-title":"Isolation of High-Purity Extracellular Vesicles by the Combination of Iodixanol Density Gradient Ultracentrifugation and Bind-Elute Chromatography From Blood Plasma","volume":"9","author":"Pelyhe","year":"2018","journal-title":"Front. Physiol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"5823","DOI":"10.1038\/s41598-021-84234-5","article-title":"Functional repertoire of EV-associated miRNA profiles after lipoprotein depletion via ultracentrifugation and size exclusion chromatography from autologous blood products","volume":"11","author":"Otahal","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_66","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_67","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1309\/AJCPRHM8CZH5EMFD","article-title":"The vascular marker CD31 also highlights histiocytes-like cells within cutaneous tumors","volume":"143","author":"Vanchinathan","year":"2015","journal-title":"Am. J. Clin. Pathol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"695","DOI":"10.3892\/ol.2016.5489","article-title":"EGFR expression is associated with cytoplasmic staining of CXCR4 and predicts poor prognosis in triple-negative breast carcinomas","volume":"13","author":"Li","year":"2017","journal-title":"Oncol. Lett."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1093\/annonc\/mdp304","article-title":"Association between gain-of-function mutations in PIK3CA and resistance to HER2-targeted agents in HER2-amplified breast cancer cell lines","volume":"21","author":"Kataoka","year":"2010","journal-title":"Ann. Oncol."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"595515","DOI":"10.3389\/fcell.2020.595515","article-title":"Hijacking Endocytosis and Autophagy in Extracellular Vesicle Communication: Where the Inside Meets the Outside","volume":"8","author":"Pedrioli","year":"2021","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"17713","DOI":"10.1074\/jbc.M112.445403","article-title":"Exosome uptake depends on ERK1\/2-heat shock protein 27 signaling and lipid Raft-mediated endocytosis negatively regulated by caveolin-1","volume":"288","author":"Svensson","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1007\/s00401-020-02254-3","article-title":"Exosomes induce endolysosomal permeabilization as a gateway by which exosomal tau seeds escape into the cytosol","volume":"141","author":"Polanco","year":"2021","journal-title":"Acta Neuropathol."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1016\/j.jconrel.2015.09.031","article-title":"Microvesicle- and exosome-mediated drug delivery enhances the cytotoxicity of Paclitaxel in autologous prostate cancer cells","volume":"220","author":"Saari","year":"2015","journal-title":"J. Control Release"},{"key":"ref_74","doi-asserted-by":"crossref","unstructured":"Antimisiaris, S.G., Mourtas, S., and Marazioti, A. (2018). Exosomes and Exosome-Inspired Vesicles for Targeted Drug Delivery. Pharmaceutics, 10.","DOI":"10.20944\/preprints201810.0507.v1"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1208\/s12248-017-0160-y","article-title":"Preservation and Storage Stability of Extracellular Vesicles for Therapeutic Applications","volume":"20","author":"Jeyaram","year":"2017","journal-title":"AAPS J."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1186\/bcr2889","article-title":"Choosing the right cell line for breast cancer research","volume":"13","author":"Holliday","year":"2011","journal-title":"Breast Cancer Res."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"3131","DOI":"10.7150\/jca.18457","article-title":"Breast Cancer Cell Line Classification and Its Relevance with Breast Tumor Subtyping","volume":"8","author":"Dai","year":"2017","journal-title":"J. Cancer"},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"4474","DOI":"10.7150\/jca.44313","article-title":"Angiogenesis in Breast Cancer Progression, Diagnosis, and Treatment","volume":"11","author":"Madu","year":"2020","journal-title":"J. Cancer"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"1282","DOI":"10.1038\/ncomms2282","article-title":"Delivery of chemotherapeutic drugs in tumour cell-derived microparticles","volume":"3","author":"Tang","year":"2012","journal-title":"Nat. Commun."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"3474","DOI":"10.7150\/thno.39434","article-title":"Tumor cell-derived exosomes home to their cells of origin and can be used as Trojan horses to deliver cancer drugs","volume":"10","author":"Qiao","year":"2020","journal-title":"Theranostics"},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"e12033","DOI":"10.1002\/jev2.12033","article-title":"Functional assays to assess the therapeutic potential of extracellular vesicles","volume":"10","author":"Nguyen","year":"2020","journal-title":"J. Extracell. Vesicles"},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.biomaterials.2018.02.030","article-title":"3D-3-culture: A tool to unveil macrophage plasticity in the tumour microenvironment","volume":"163","author":"Rebelo","year":"2018","journal-title":"Biomaterials"},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"e12085","DOI":"10.1002\/jev2.12085","article-title":"Biodistribution of extracellular vesicles following administration into animals: A systematic review","volume":"10","author":"Kang","year":"2021","journal-title":"J. Extracell. Vesicles"}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/15\/3\/953\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:55:41Z","timestamp":1760122541000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/15\/3\/953"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,15]]},"references-count":83,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2023,3]]}},"alternative-id":["pharmaceutics15030953"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics15030953","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,15]]}}}