{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T12:54:51Z","timestamp":1782824091756,"version":"3.54.5"},"posted":{"date-parts":[[2024,1,16]]},"group-title":"Bioengineering","reference-count":66,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,7,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Ischemic stroke-induced mitochondrial dysfunction in the blood-brain barrier-forming brain endothelial cells (\n                  <jats:bold>BECs<\/jats:bold>\n                  ) results in long-term neurological dysfunction post-stroke. We previously data from a pilot study where\n                  <jats:italic>intravenous<\/jats:italic>\n                  administration of human BEC (\n                  <jats:bold>hBEC<\/jats:bold>\n                  )-derived mitochondria-containing extracellular vesicles (\n                  <jats:bold>EVs<\/jats:bold>\n                  ) showed a potential efficacy signal in a mouse middle cerebral artery occlusion (\n                  <jats:bold>MCAo<\/jats:bold>\n                  ) model of stroke. We\n                  <jats:italic>hypothesized<\/jats:italic>\n                  that EVs harvested from donor species homologous to the recipient species (\n                  <jats:italic>e.g.,<\/jats:italic>\n                  mouse) may improve therapeutic efficacy, and therefore, use of mouse BEC (\n                  <jats:bold>mBEC<\/jats:bold>\n                  )-derived EVs may improve post-stroke outcomes in MCAo mice.\n                <\/jats:p>\n                <jats:p>\n                  We investigated potential differences in the mitochondria transfer of EVs derived from the same species as the recipient cell (mBEC-EVs and recipient mBECs or hBECs-EVs and recipient hBECs)\n                  <jats:italic>vs<\/jats:italic>\n                  . cross-species EVs and recipient cells (mBEC-EVs and recipient hBECs or\n                  <jats:italic>vice versa<\/jats:italic>\n                  ). Our results showed that while both hBEC- and mBEC-EVs transferred EV mitochondria, mBEC-EVs outperformed hBEC-EVs in increasing ATP levels and improved recipient mBEC mitochondrial function via increasing oxygen consumption rates. mBEC-EVs significantly reduced brain infarct volume and neurological deficit scores compared to vehicle-injected MCAo mice. The superior therapeutic efficacy of mBEC-EVs in a mouse MCAo stroke support the continued use of mBEC-EVs to optimize the therapeutic potential of mitochondria-containing EVs in preclinical mouse models.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Graphical Abstract<\/jats:title>\n                  <jats:fig id=\"ufig1\" position=\"float\" orientation=\"portrait\" fig-type=\"figure\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"575903v2_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:fig>\n                <\/jats:sec>","DOI":"10.1101\/2024.01.16.575903","type":"posted-content","created":{"date-parts":[[2024,1,17]],"date-time":"2024-01-17T03:10:10Z","timestamp":1705461010000},"source":"Crossref","is-referenced-by-count":3,"title":["Mitochondria-containing extracellular vesicles from mouse\n                  <i>vs<\/i>\n                  . human brain endothelial cells for ischemic stroke therapy"],"prefix":"10.64898","author":[{"given":"Kandarp M.","family":"Dave","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Venugopal R.","family":"Venna","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Krithika S.","family":"Rao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Donna B.","family":"Stolz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bodhi","family":"Brady","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victoria A.","family":"Quaicoe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael E.","family":"Maniskas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ella E.","family":"Hildebrand","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dawson","family":"Green","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingxi","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jadranka","family":"Milosevic","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Si-yang","family":"Zheng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sruti S.","family":"Shiva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Louise D.","family":"McCullough","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Devika S","family":"Manickam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2024071304450467000_2024.01.16.575903v2.1","doi-asserted-by":"publisher","DOI":"10.14336\/AD.2017.1126"},{"key":"2024071304450467000_2024.01.16.575903v2.2","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1016\/j.redox.2018.03.002","article-title":"Diverse roles of mitochondria in ischemic stroke","volume":"16","year":"2018","journal-title":"Redox Biol"},{"key":"2024071304450467000_2024.01.16.575903v2.3","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.111.300233"},{"issue":"1","key":"2024071304450467000_2024.01.16.575903v2.4","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1186\/s12967-023-04493-w","article-title":"Mitochondrial dysfunction in neurodegenerative disorders: Potential therapeutic application of mitochondrial transfer to central nervous system-residing cells","volume":"21","year":"2023","journal-title":"Journal of Translational Medicine"},{"key":"2024071304450467000_2024.01.16.575903v2.5","doi-asserted-by":"publisher","DOI":"10.1161\/CIR.0000000000000485"},{"key":"2024071304450467000_2024.01.16.575903v2.6","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.00095.2018"},{"key":"2024071304450467000_2024.01.16.575903v2.7","doi-asserted-by":"crossref","unstructured":"Nian K , Harding IC , Herman IM , Ebong EE. Blood-Brain Barrier Damage in Ischemic Stroke and Its Regulation by Endothelial Mechanotransduction. Frontiers in Physiology. 2020;11(1681).","DOI":"10.3389\/fphys.2020.605398"},{"key":"2024071304450467000_2024.01.16.575903v2.8","doi-asserted-by":"publisher","DOI":"10.1002\/ana.410010502"},{"issue":"7","key":"2024071304450467000_2024.01.16.575903v2.9","doi-asserted-by":"crossref","first-page":"1546","DOI":"10.1177\/0271678X19900030","article-title":"Endothelial-specific Crif1 deletion induces BBB maturation and disruption via the alteration of actin dynamics by impaired mitochondrial respiration","volume":"40","year":"2020","journal-title":"J Cereb Blood Flow Metab"},{"key":"2024071304450467000_2024.01.16.575903v2.10","doi-asserted-by":"crossref","unstructured":"Bernardo-Castro S , Sousa JA , Br\u00e1s A , Cec\u00edlia C , Rodrigues B , Almendra L , Machado C , Santo G , Silva F , Ferreira L , Santana I , Sargento-Freitas J. Pathophysiology of Blood\u2013Brain Barrier Permeability Throughout the Different Stages of Ischemic Stroke and Its Implication on Hemorrhagic Transformation and Recovery. Frontiers in Neurology. 2020;11(1605).","DOI":"10.3389\/fneur.2020.594672"},{"issue":"1","key":"2024071304450467000_2024.01.16.575903v2.11","doi-asserted-by":"crossref","first-page":"15717","DOI":"10.1038\/s41598-019-52083-y","article-title":"Mitochondria damaged by Oxygen Glucose Deprivation can be Restored through Activation of the PI3K\/Akt Pathway and Inhibition of Calcium Influx by Amlodipine Camsylate","volume":"9","year":"2019","journal-title":"Scientific Reports"},{"key":"2024071304450467000_2024.01.16.575903v2.12","doi-asserted-by":"publisher","DOI":"10.1002\/jcb.20837"},{"key":"2024071304450467000_2024.01.16.575903v2.13","doi-asserted-by":"publisher","DOI":"10.1038\/sj.emboj.7601963"},{"key":"2024071304450467000_2024.01.16.575903v2.14","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1016\/j.jconrel.2022.01.045","article-title":"Delivery of mitochondria via extracellular vesicles \u2013 A new horizon in drug delivery","volume":"343","year":"2022","journal-title":"Journal of Controlled Release"},{"issue":"1","key":"2024071304450467000_2024.01.16.575903v2.15","doi-asserted-by":"crossref","first-page":"1971","DOI":"10.1038\/s41467-021-21984-w","article-title":"Selective packaging of mitochondrial proteins into extracellular vesicles prevents the release of mitochondrial DAMPs","volume":"12","year":"2021","journal-title":"Nature Communications"},{"issue":"1","key":"2024071304450467000_2024.01.16.575903v2.16","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s00702-009-0288-8","article-title":"Astrocytes and Glioblastoma cells release exosomes carrying mtDNA","volume":"117","year":"2010","journal-title":"J Neural Transm (Vienna"},{"key":"2024071304450467000_2024.01.16.575903v2.17","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.118.314601"},{"key":"2024071304450467000_2024.01.16.575903v2.18","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms9472"},{"key":"2024071304450467000_2024.01.16.575903v2.19","doi-asserted-by":"publisher","DOI":"10.1016\/j.jacc.2020.12.060"},{"key":"2024071304450467000_2024.01.16.575903v2.20","doi-asserted-by":"crossref","unstructured":"Banks WA , Sharma P , Bullock KM , Hansen KM , Ludwig N , Whiteside TL. Transport of Extracellular Vesicles across the Blood-Brain Barrier: Brain Pharmacokinetics and Effects of Inflammation. Int J Mol Sci. 2020;21(12).","DOI":"10.3390\/ijms21124407"},{"key":"2024071304450467000_2024.01.16.575903v2.21","doi-asserted-by":"publisher","DOI":"10.1038\/s12276-019-0280-9"},{"key":"2024071304450467000_2024.01.16.575903v2.22","doi-asserted-by":"publisher","DOI":"10.1016\/j.jconrel.2017.07.001"},{"key":"2024071304450467000_2024.01.16.575903v2.23","doi-asserted-by":"crossref","first-page":"1037","DOI":"10.1016\/j.jconrel.2023.12.021","article-title":"Simultaneous ischemic regions targeting and BBB crossing strategy to harness extracellular vesicles for therapeutic delivery in ischemic stroke","volume":"365","year":"2024","journal-title":"Journal of Controlled Release"},{"key":"2024071304450467000_2024.01.16.575903v2.24","doi-asserted-by":"crossref","first-page":"32570","DOI":"10.3402\/jev.v5.32570","article-title":"High-resolution proteomic and lipidomic analysis of exosomes and microvesicles from different cell sources","volume":"5","year":"2016","journal-title":"Journal of extracellular vesicles"},{"issue":"1","key":"2024071304450467000_2024.01.16.575903v2.25","doi-asserted-by":"crossref","first-page":"1302705","DOI":"10.1080\/20013078.2017.1302705","article-title":"Characterisation of extracellular vesicle-subsets derived from brain endothelial cells and analysis of their protein cargo modulation after TNF exposure","volume":"6","year":"2017","journal-title":"Journal of Extracellular Vesicles"},{"key":"2024071304450467000_2024.01.16.575903v2.26","doi-asserted-by":"publisher","DOI":"10.1080\/21541248.2016.1215283"},{"issue":"1","key":"2024071304450467000_2024.01.16.575903v2.27","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","year":"2018","journal-title":"J Extracell Vesicles"},{"key":"2024071304450467000_2024.01.16.575903v2.28","doi-asserted-by":"publisher","DOI":"10.1002\/jev2.12404"},{"key":"2024071304450467000_2024.01.16.575903v2.29","doi-asserted-by":"publisher","DOI":"10.1038\/nm.2736"},{"key":"2024071304450467000_2024.01.16.575903v2.30","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.redox.2018.06.009","article-title":"Exosomal transfer of mitochondria from airway myeloid-derived regulatory cells to T cells","volume":"18","year":"2018","journal-title":"Redox Biol"},{"key":"2024071304450467000_2024.01.16.575903v2.31","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1016\/j.mito.2020.08.005","article-title":"The existence and function of mitochondrial component in extracellular vesicles","volume":"54","year":"2020","journal-title":"Mitochondrion"},{"key":"2024071304450467000_2024.01.16.575903v2.32","doi-asserted-by":"publisher","DOI":"10.1183\/13993003.02978-2020"},{"key":"2024071304450467000_2024.01.16.575903v2.33","doi-asserted-by":"publisher","DOI":"10.1016\/j.jconrel.2023.01.025"},{"key":"2024071304450467000_2024.01.16.575903v2.34","doi-asserted-by":"publisher","DOI":"10.1016\/j.jconrel.2021.08.038"},{"key":"2024071304450467000_2024.01.16.575903v2.35","doi-asserted-by":"crossref","unstructured":"Dave KM , Stolz DB , Manickam DS. Delivery of mitochondria-containing extracellular vesicles to the BBB for ischemic stroke therapy. Expert Opinion on Drug Delivery.1\u201320.","DOI":"10.1080\/17425247.2023.2279115"},{"issue":"1","key":"2024071304450467000_2024.01.16.575903v2.36","doi-asserted-by":"crossref","first-page":"1324730","DOI":"10.1080\/20013078.2017.1324730","article-title":"Comprehensive toxicity and immunogenicity studies reveal minimal effects in mice following sustained dosing of extracellular vesicles derived from HEK293T cells","volume":"6","year":"2017","journal-title":"J Extracell Vesicles"},{"key":"2024071304450467000_2024.01.16.575903v2.37","doi-asserted-by":"publisher","DOI":"10.1208\/s12249-020-01892-w"},{"issue":"5","key":"2024071304450467000_2024.01.16.575903v2.38","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1007\/s12195-022-00738-8","article-title":"Engineering Extracellular Vesicles to Modulate Their Innate Mitochondrial Load","volume":"15","year":"2022","journal-title":"Cell Mol Bioeng"},{"issue":"3","key":"2024071304450467000_2024.01.16.575903v2.39","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1177\/0271678X19830791","article-title":"TGF\u03b1 preserves oligodendrocyte lineage cells and improves white matter integrity after cerebral ischemia","volume":"40","year":"2020","journal-title":"Journal of Cerebral Blood Flow & Metabolism"},{"key":"2024071304450467000_2024.01.16.575903v2.40","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2013-09-529420"},{"key":"2024071304450467000_2024.01.16.575903v2.41","doi-asserted-by":"crossref","unstructured":"McCullough LD , Roy-O\u2019Reilly M , Lai Y-J , Patrizz A , Xu Y , Lee J , Holmes A , Kraushaar DC , Chauhan A , Sansing LH , Stonestreet BS , Zhu L , Kofler J , Lim Y-P , Venna VR. Exogenous inter-\u03b1 inhibitor proteins prevent cell death and improve ischemic stroke outcomes in mice. The Journal of Clinical Investigation. 2021;131(17).","DOI":"10.1172\/JCI144898"},{"issue":"4","key":"2024071304450467000_2024.01.16.575903v2.42","doi-asserted-by":"crossref","first-page":"792","DOI":"10.1038\/jcbfm.2009.5","article-title":"Changes in Experimental Stroke Outcome across the Life Span","volume":"29","year":"2009","journal-title":"Journal of Cerebral Blood Flow & Metabolism"},{"key":"2024071304450467000_2024.01.16.575903v2.43","doi-asserted-by":"publisher","DOI":"10.1007\/s00401-012-0990-8"},{"key":"2024071304450467000_2024.01.16.575903v2.44","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M409985200"},{"key":"2024071304450467000_2024.01.16.575903v2.45","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1607002114"},{"key":"2024071304450467000_2024.01.16.575903v2.46","first-page":"575","article-title":"Early retinal inflammatory biomarkers in the middle cerebral artery occlusion model of ischemic stroke","volume":"22","year":"2016","journal-title":"Mol Vis"},{"key":"2024071304450467000_2024.01.16.575903v2.47","doi-asserted-by":"publisher","DOI":"10.1101\/lm.038083.115"},{"key":"2024071304450467000_2024.01.16.575903v2.48","doi-asserted-by":"crossref","unstructured":"Pospichalova V , Svoboda J , Dave Z , Kotrbova A , Kaiser K , Klemova D , Ilkovics L , Hampl A , Crha I , Jandakova E , Minar L , Weinberger V , Bryja V. Simplified protocol for flow cytometry analysis of fluorescently labeled exosomes and microvesicles using dedicated flow cytometer. J Extracell Vesicles. 2015;4:25530.","DOI":"10.3402\/jev.v4.25530"},{"issue":"3","key":"2024071304450467000_2024.01.16.575903v2.49","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1016\/j.jaccao.2021.05.006","article-title":"Mitochondria-Rich Extracellular Vesicles Rescue Patient-Specific Cardiomyocytes From Doxorubicin Injury: Insights Into the SENECA Trial","volume":"3","year":"2021","journal-title":"JACC CardioOncol"},{"key":"2024071304450467000_2024.01.16.575903v2.50","doi-asserted-by":"publisher","DOI":"10.1038\/nature18928"},{"issue":"8","key":"2024071304450467000_2024.01.16.575903v2.51","doi-asserted-by":"crossref","first-page":"3684","DOI":"10.7150\/thno.41580","article-title":"Progress, opportunity, and perspective on exosome isolation - efforts for efficient exosome-based theranostics","volume":"10","year":"2020","journal-title":"Theranostics"},{"issue":"1","key":"2024071304450467000_2024.01.16.575903v2.52","doi-asserted-by":"crossref","first-page":"17319","DOI":"10.1038\/srep17319","article-title":"Isolation of exosomes by differential centrifugation: Theoretical analysis of a commonly used protocol","volume":"5","year":"2015","journal-title":"Scientific Reports"},{"key":"2024071304450467000_2024.01.16.575903v2.53","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1418401112"},{"key":"2024071304450467000_2024.01.16.575903v2.54","doi-asserted-by":"crossref","unstructured":"Konoshenko MY , Lekchnov EA , Vlassov AV , Laktionov PP. Isolation of Extracellular Vesicles: General Methodologies and Latest Trends. Biomed Res Int. 2018;2018:8545347.","DOI":"10.1155\/2018\/8545347"},{"key":"2024071304450467000_2024.01.16.575903v2.55","doi-asserted-by":"publisher","DOI":"10.1038\/nrd3978"},{"issue":"2","key":"2024071304450467000_2024.01.16.575903v2.56","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1002\/cyto.a.22795","article-title":"Analytical challenges of extracellular vesicle detection: A comparison of different techniques","volume":"89","year":"2016","journal-title":"Cytometry Part A"},{"key":"2024071304450467000_2024.01.16.575903v2.57","doi-asserted-by":"crossref","unstructured":"Andreu Z , Y\u00e1\u00f1ez-M\u00f3 M. Tetraspanins in extracellular vesicle formation and function. Front Immunol. 2014;5:442.","DOI":"10.3389\/fimmu.2014.00442"},{"key":"2024071304450467000_2024.01.16.575903v2.58","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2009.09.059"},{"issue":"9","key":"2024071304450467000_2024.01.16.575903v2.59","doi-asserted-by":"crossref","first-page":"1404","DOI":"10.1002\/stem.2856","article-title":"Protective Effects of Endothelial Progenitor Cell-Derived Extracellular Mitochondria in Brain Endothelium","volume":"36","year":"2018","journal-title":"Stem Cells"},{"key":"2024071304450467000_2024.01.16.575903v2.60","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1014918108"},{"key":"2024071304450467000_2024.01.16.575903v2.61","unstructured":"A. Naveena H , Bhatia D. Hypoxia Modulates Cellular Endocytic Pathways and Organelles with Enhanced Cell Migration and 3D Cell Invasion**. ChemBioChem.n\/a(n\/a):e202300506."},{"issue":"1","key":"2024071304450467000_2024.01.16.575903v2.62","doi-asserted-by":"crossref","first-page":"37959","DOI":"10.1038\/srep37959","article-title":"TDB protects vascular endothelial cells against oxygen-glucose deprivation\/reperfusion-induced injury by targeting miR-34a to increase Bcl-2 expression","volume":"6","year":"2016","journal-title":"Scientific Reports"},{"key":"2024071304450467000_2024.01.16.575903v2.63","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1007\/978-1-4939-6960-9_7","article-title":"Assaying Mitochondrial Respiration as an Indicator of Cellular Metabolism and Fitness","volume":"1601","year":"2017","journal-title":"Methods Mol Biol"},{"key":"2024071304450467000_2024.01.16.575903v2.64","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0085436"},{"key":"2024071304450467000_2024.01.16.575903v2.65","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0231267"},{"issue":"3","key":"2024071304450467000_2024.01.16.575903v2.66","doi-asserted-by":"crossref","first-page":"100508","DOI":"10.1016\/j.xpro.2021.100508","article-title":"Protocols for endothelial cell isolation from mouse tissues: brain, choroid, lung, and muscle","volume":"2","year":"2021","journal-title":"STAR Protocols"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2024.01.16.575903","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T05:48:20Z","timestamp":1768456100000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2024.01.16.575903"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,1,16]]},"references-count":66,"URL":"https:\/\/doi.org\/10.1101\/2024.01.16.575903","relation":{},"subject":[],"published":{"date-parts":[[2024,1,16]]},"subtype":"preprint"}}