{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,7]],"date-time":"2025-10-07T11:46:52Z","timestamp":1759837612825,"version":"3.41.2"},"reference-count":35,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,6,16]],"date-time":"2023-06-16T00:00:00Z","timestamp":1686873600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["GNT1163282","GNT2003156"],"award-info":[{"award-number":["GNT1163282","GNT2003156"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Neurosci."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Thrombolytic agents such as tissue plasminogen activator (tPA) are the only drug class approved to treat ischemic stroke and are usually administered within 4.5\u2009h. However, only ~20% of ischemic stroke patients are eligible to receive the therapy. We previously demonstrated that early intravenous administration of human amnion epithelial cells (hAECs) can limit brain inflammation and infarct growth in experimental stroke. Here, we have tested whether hAECs exert cerebroprotective effects in combination with tPA in mice.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Male C57Bl\/6 mice were subjected to middle cerebral artery occlusion for 60\u2009min followed by reperfusion. Immediately following reperfusion, vehicle (saline, <jats:italic>n<\/jats:italic>\u2009=\u200931) or tPA (10\u2009mg\/kg; <jats:italic>n<\/jats:italic>\u2009=\u200973) was administered intravenously. After 30\u2009min of reperfusion, tPA-treated mice were injected intravenously with either hAECs (1\u00d710<jats:sup>6<\/jats:sup>; <jats:italic>n<\/jats:italic>\u2009=\u200932) or vehicle (2% human serum albumin; <jats:italic>n<\/jats:italic>\u2009=\u200941). A further 15 sham-operated mice were treated with vehicle (<jats:italic>n<\/jats:italic>\u2009=\u20097) or tPA\u2009+\u2009vehicle (<jats:italic>n<\/jats:italic>\u2009=\u20098). Mice were designated to be euthanised at 3, 6 or 24\u2009h post-stroke (<jats:italic>n<\/jats:italic>\u2009=\u200921, 31, and 52, respectively), and brains were collected to assess infarct volume, blood\u2013brain barrier (BBB) disruption, intracerebral bleeding and inflammatory cell content.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>There was no mortality within 6\u2009h of stroke onset, but a high mortality occurred in tPA\u2009+\u2009saline-treated mice between 6\u2009h and 24\u2009h post-stroke in comparison to mice treated with tPA\u2009+\u2009hAECs (61% vs. 27%, <jats:italic>p<\/jats:italic>\u2009=\u20090.04). No mortality occurred within 24\u2009h of sham surgery in mice treated with tPA\u2009+\u2009vehicle. We focused on early infarct expansion within 6\u2009h of stroke and found that infarction was ~50% larger in tPA\u2009+\u2009saline- than in vehicle-treated mice (23\u2009\u00b1\u20093\u2009mm<jats:sup>3<\/jats:sup> vs. 15\u2009\u00b1\u20092\u2009mm<jats:sup>3<\/jats:sup>, <jats:italic>p<\/jats:italic>\u2009=\u20090.02) but not in mice receiving tPA\u2009+\u2009hAECs (13\u2009\u00b1\u20092\u2009mm<jats:sup>3<\/jats:sup>, <jats:italic>p<\/jats:italic>\u2009&amp;lt;\u20090.01 vs. tPA\u2009+\u2009saline) in which intracerebral hAECs were detected. Similar to the profiles of infarct expansion, BBB disruption and intracerebral bleeding in tPA\u2009+\u2009saline-treated mice at 6\u2009h was 50\u201360% greater than in vehicle-treated controls (2.6\u2009\u00b1\u20090.5 vs. 1.6\u2009\u00b1\u20090.2, <jats:italic>p<\/jats:italic>\u2009=\u20090.05) but not after tPA\u2009+\u2009hAECs treatment (1.7\u2009\u00b1\u20090.2, <jats:italic>p<\/jats:italic>\u2009=\u20090.10 vs. tPA\u2009+\u2009saline). No differences in inflammatory cell content were detected between treatment groups.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>When administered following tPA in acute stroke, hAECs improve safety and attenuate infarct growth in association with less BBB disruption and lower 24\u2009h mortality.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fnins.2023.1157236","type":"journal-article","created":{"date-parts":[[2023,6,16]],"date-time":"2023-06-16T04:42:23Z","timestamp":1686890543000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":4,"title":["Protection against brain injury after ischemic stroke by intravenous human amnion epithelial cells in combination with tissue plasminogen activator"],"prefix":"10.3389","volume":"17","author":[{"given":"Liz J.","family":"Barreto-Arce","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hyun Ah","family":"Kim","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Siow Teng","family":"Chan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rebecca","family":"Lim","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Grant R.","family":"Drummond","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Henry","family":"Ma","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thanh G.","family":"Phan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher G.","family":"Sobey","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shenpeng R.","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2023,6,16]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"243","DOI":"10.2174\/187152708784936608","article-title":"Tissue plasminogen activator (tPA) and matrix metalloproteinases in the pathogenesis of stroke: therapeutic strategies","volume":"7","author":"Adibhatla","year":"2008","journal-title":"CNS Neurol. Disord. Drug Targets"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"113275","DOI":"10.1016\/j.expneurol.2020.113275","article-title":"Human neural stem cells improve early stage stroke outcome in delayed tissue plasminogen activator-treated aged stroke brains","volume":"329","author":"Boese","year":"2020","journal-title":"Exp. Neurol."},{"key":"ref140","doi-asserted-by":"publisher","first-page":"66","DOI":"10.3389\/fncel.2012.00066","article-title":"Post-stroke inflammation and the potential efficacy of novel stem cell therapies: focus on amnion epithelial cells","volume":"6","author":"Broughton","year":"2013","journal-title":"Front. Cell. Neurosci"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"70","DOI":"10.1038\/s41572-019-0118-8","article-title":"Ischaemic stroke","volume":"5","author":"Campbell","year":"2019","journal-title":"Nat. Rev. Dis. Primers."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"194","DOI":"10.3389\/fnins.2013.00194","article-title":"Stem cell therapy to protect and repair the developing brain: a review of mechanisms of action of cord blood and amnion epithelial derived cells","volume":"7","author":"Castillo-Melendez","year":"2013","journal-title":"Front. Neurosci."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1186\/s12974-018-1153-1","article-title":"Mesenchymal stem cells attenuate blood-barrier leakage after cerebral ischemia in mice","volume":"15","author":"Cheng","year":"2018","journal-title":"J. Neuroinflammation"},{"key":"ref130","first-page":"11","article-title":"The cerebral embolism evoked by intra-arterial delivery of allogeneic bone marrow mesenchymal stem cells in rats is related to cell dose and infusion velocity","volume":"6","author":"Cui","year":"2015","journal-title":"Stem Cell Res. The"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"700","DOI":"10.1161\/STROKEAHA.117.019136","article-title":"Acute or delayed systemic Administration of Human Amnion Epithelial Cells Improves Outcomes in experimental stroke","volume":"49","author":"Evans","year":"2018","journal-title":"Stroke"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"972","DOI":"10.1016\/j.neuropharm.2009.12.017","article-title":"Blood-brain barrier permeability and tPA-mediated neurotoxicity","volume":"58","author":"Fanne","year":"2010","journal-title":"Neuropharmacology"},{"key":"ref120","doi-asserted-by":"publisher","first-page":"2244","DOI":"10.1161\/STROKEAHA.108.541128","article-title":"Update of the stroke therapy academic industry roundtable preclinical recommendations","volume":"40","author":"Fisher","year":"2009","journal-title":"Stroke"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1517\/14728222.12.2.159","article-title":"Tissue-type plasminogen activator as a therapeutic target in stroke","volume":"12","author":"Gravanis","year":"2008","journal-title":"Expert Opin. Ther. Targets"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"1804","DOI":"10.1038\/jcbfm.2010.149","article-title":"The contribution of L-arginine to the neurotoxicity of recombinant tissue plasminogen activator following cerebral ischemia: a review of rtPA neurotoxicity","volume":"30","author":"Harston","year":"2010","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"ref10","doi-asserted-by":"publisher","first-page":"680","DOI":"10.1111\/j.1476-5381.2008.00073.x","article-title":"Reduction of cerebral infarct volume by apocynin requires pretreatment and is absent in Nox2-deficient mice","volume":"156","author":"Jackman","year":"2009","journal-title":"Br. J. Pharmacol."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"3310","DOI":"10.1161\/STROKEAHA.115.008861","article-title":"Assessment of blood\u2013brain barrier disruption in stroke","volume":"46","author":"Kassner","year":"2015","journal-title":"Stroke"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"2641","DOI":"10.1161\/hs1101.097381","article-title":"Effects of recombinant tissue plasminogen activator after intraluminal thread occlusion in mice: role of hemodynamic alterations","volume":"32","author":"Kilic","year":"2001","journal-title":"Stroke"},{"key":"ref13","doi-asserted-by":"publisher","first-page":"160","DOI":"10.5853\/jos.2019.00584","article-title":"tPA helpers in the treatment of acute ischemic stroke: are they ready for clinical use?","volume":"21","author":"Kim","year":"2019","journal-title":"J. Stroke"},{"key":"ref14","doi-asserted-by":"publisher","first-page":"100072","DOI":"10.1016\/j.bbih.2020.100072","article-title":"Systemic treatment with human amnion epithelial cells after experimental traumatic brain injury","volume":"5","author":"Kim","year":"2020","journal-title":"Brain, Behav. Immunity-Health"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"365","DOI":"10.3389\/fncel.2014.00365","article-title":"Brain immune cell composition and functional outcome after cerebral ischemia: comparison of two mouse strains","volume":"8","author":"Kim","year":"2014","journal-title":"Front. Cell. Neurosci."},{"key":"ref16","doi-asserted-by":"publisher","first-page":"561","DOI":"10.1055\/s-0038-1653411","article-title":"Studies on the specific fibrinolytic effect of human extrinsic (tissue-type) plasminogen activator in human blood and in various animal species in vitro","volume":"46","author":"Korninger","year":"1981","journal-title":"Thromb. Haemost."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"656","DOI":"10.3389\/fneur.2019.00656","article-title":"Cell-based therapies for stroke: are we there yet?","volume":"10","author":"Krause","year":"2019","journal-title":"Front. Neurol."},{"key":"ref18","doi-asserted-by":"publisher","first-page":"2086","DOI":"10.1002\/jnr.23104","article-title":"Intracerebral transplantation of bone marrow stromal cells ameliorates tissue plasminogen activator-induced brain damage after cerebral ischemia in mice detected by in vivo and ex vivo optical imaging","volume":"90","author":"Liu","year":"2012","journal-title":"J. Neurosci. Res."},{"key":"ref19","doi-asserted-by":"publisher","first-page":"1795","DOI":"10.1056\/NEJMoa1813046","article-title":"Thrombolysis guided by perfusion imaging up to 9 hours after onset of stroke","volume":"380","author":"Ma","year":"2019","journal-title":"N. Engl. J. Med."},{"key":"ref20","doi-asserted-by":"publisher","first-page":"1549","DOI":"10.1634\/stemcells.2004-0357","article-title":"Stem cell characteristics of amniotic epithelial cells","volume":"23","author":"Miki","year":"2005","journal-title":"Stem Cells (Dayton, Ohio)"},{"key":"ref21","doi-asserted-by":"publisher","first-page":"Unit 1E.6","DOI":"10.1002\/9780470151808.sc01e06s13","article-title":"Amnion epithelial cell isolation and characterization for clinical use","author":"Murphy","year":"2010","journal-title":"Curr. Protoc. Stem Cell Biol."},{"key":"ref22","doi-asserted-by":"publisher","first-page":"917","DOI":"10.3171\/2016.8.JNS16240","article-title":"Intravenous infusion of mesenchymal stem cells inhibits intracranial hemorrhage after recombinant tissue plasminogen activator therapy for transient middle cerebral artery occlusion in rats","volume":"127","author":"Nakazaki","year":"2017","journal-title":"J. Neurosurg."},{"key":"ref100","doi-asserted-by":"publisher","first-page":"1153231","DOI":"10.3389\/fnins.2023.1153231","article-title":"Phase I trial outcome of amnion cell therapy in patients with ischemic stroke (I-ACT)","volume":"17","author":"Phan","year":"2023","journal-title":"Front Neurosci"},{"volume-title":"National Stroke Audit Acute Services Report","year":"2017","key":"ref23"},{"key":"ref24","doi-asserted-by":"publisher","first-page":"1844","DOI":"10.1161\/STROKEAHA.115.012577","article-title":"Eligibility and predictors for acute revascularization procedures in a stroke center","volume":"47","author":"Vanacker","year":"2016","journal-title":"Stroke"},{"key":"ref25","doi-asserted-by":"publisher","first-page":"566","DOI":"10.1016\/j.neurol.2017.07.003","article-title":"Can the benefits of rtPA treatment for stroke be improved?","volume":"173","author":"Vivien","year":"2017","journal-title":"Rev. Neurol."},{"key":"ref26","doi-asserted-by":"publisher","first-page":"228","DOI":"10.1038\/nm0298-228","article-title":"Tissue plasminogen activator (tPA) increases neuronal damage after focal cerebral ischemia in wild-type and tPA-deficient mice","volume":"4","author":"Wang","year":"1998","journal-title":"Nat. Med."},{"key":"ref27","doi-asserted-by":"publisher","first-page":"4380","DOI":"10.3390\/ijms22094380","article-title":"Plasminogen activators in neurovascular and neurodegenerative disorders","volume":"22","author":"Yepes","year":"2021","journal-title":"Int. J. Mol. Sci."},{"key":"ref28","doi-asserted-by":"publisher","first-page":"286","DOI":"10.5853\/jos.2019.03048","article-title":"Optimizing stem cell therapy after ischemic brain injury","volume":"22","author":"Zhang","year":"2020","journal-title":"J. Stroke"},{"key":"ref29","doi-asserted-by":"publisher","first-page":"439","DOI":"10.1186\/s13287-020-01951-w","article-title":"Application of human amniotic epithelial cells in regenerative medicine: a systematic review","volume":"11","author":"Zhang","year":"2020","journal-title":"Stem Cell Res Ther"},{"key":"ref30","doi-asserted-by":"publisher","first-page":"96","DOI":"10.1016\/j.tips.2020.11.010","article-title":"Targeting the immune system for ischemic stroke","volume":"42","author":"Zhang","year":"2021","journal-title":"Trends Pharmacol. Sci."},{"key":"ref31","doi-asserted-by":"publisher","first-page":"1489","DOI":"10.1007\/s00018-019-03005-8","article-title":"The role of endogenous tissue-type plasminogen activator in neuronal survival after ischemic stroke: friend or foe?","volume":"76","author":"Zhu","year":"2019","journal-title":"Cell. Mol. Life Sci."}],"container-title":["Frontiers in Neuroscience"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fnins.2023.1157236\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,6,16]],"date-time":"2023-06-16T04:42:27Z","timestamp":1686890547000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fnins.2023.1157236\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,6,16]]},"references-count":35,"alternative-id":["10.3389\/fnins.2023.1157236"],"URL":"https:\/\/doi.org\/10.3389\/fnins.2023.1157236","relation":{},"ISSN":["1662-453X"],"issn-type":[{"type":"electronic","value":"1662-453X"}],"subject":[],"published":{"date-parts":[[2023,6,16]]},"article-number":"1157236"}}