Name: ferric chloride-induced murine thrombosis models
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Description: Watch this Scientific Journal Video about Ferric Chloride-induced Murine Thrombosis Models at JoVE.com

This work was supported by the National Heart Lung and Blood Institute (NHLBI) of the National Institutes of Health under award numbers R01 HL121212 (PI: Sen Gupta), R01 HL129179 (PI: Sen Gupta, Co-I: Li) and R01 HL098217 (PI: Nieman). The content of this publication is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Alla förfaranden och manipulationer av djur har godkänts av Institutional Animal Care och användning kommittéer (IACUC) The Cleveland Clinic i enlighet med United States Public Health Service politik om human vård och användning av djur, och NIH Guide för vård och användningen av försöksdjur. 1. Förberedelser: <ol><li> Fluorescerande färgämne för Märkning Trombocyter <ol><li> Förbereda rodamin 6G-lösning, 0,5 mg / ml, i saltlösning och sterilisera lösninge...

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	<li>Sachs, U. J., Nieswandt, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+thrombus+formation+in+murine+models.">In vivo thrombus formation in murine models.</a> Circ Res. 100, 979-991 (2007).</li><li>Libby, P., Lichtman, A. H., Hansson, G. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immune+effector+mechanisms+implicated+in+atherosclerosis:+from+mice+to+humans.">Immune effector mechanisms implicated in atherosclerosis: from mice to humans.</a> Immunity. 38, 1092-1104 (2013).</li><li>Ruggeri, Z. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Platelet+adhesion+under+flow.">Platelet adhesion under flow.</a> Microcirculation. 16, 58-83 (2009).</li><li>Watson, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Platelet+activation+by+extracellular+matrix+proteins+in+haemostasis+and+thrombosis.">Platelet activation by extracellular matrix proteins in haemostasis and thrombosis.</a> Curr Pharm Des. 15, 1358-1372 (2009).</li><li>Furie, B., Furie, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thrombus+formation+in+vivo.">Thrombus formation in vivo.</a> J Clin Invest. 115, 3355-3362 (2005).</li><li>Li, W., McIntyre, T. M., Silverstein, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ferric+chloride-induced+murine+carotid+arterial+injury:+A+model+of+redox+pathology.">Ferric chloride-induced murine carotid arterial injury: A model of redox pathology.</a> Redox Biol. 1, 50-55 (2013).</li><li>Ghosh, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Platelet+CD36+mediates+interactions+with+endothelial+cell-derived+microparticles+and+contributes+to+thrombosis+in+mice.">Platelet CD36 mediates interactions with endothelial cell-derived microparticles and contributes to thrombosis in mice.</a> J Clin Invest. 118, 1934-1943 (2008).</li><li>Chen, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vav+guanine+nucleotide+exchange+factors+link+hyperlipidemia+and+a+prothrombotic+state.">Vav guanine nucleotide exchange factors link hyperlipidemia and a prothrombotic state.</a> Blood. , (2011).</li><li>Li, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CD36+participates+in+a+signaling+pathway+that+regulates+ROS+formation+in+murine+VSMCs.">CD36 participates in a signaling pathway that regulates ROS formation in murine VSMCs.</a> J Clin Invest. 120, 3996-4006 (2010).</li><li>Chen, K., Febbraio, M., Li, W., Silverstein, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+specific+CD36-dependent+signaling+pathway+is+required+for+platelet+activation+by+oxidized+low-density+lipoprotein.">A specific CD36-dependent signaling pathway is required for platelet activation by oxidized low-density lipoprotein.</a> Circ Res. 102, 1512-1519 (2008).</li><li>Kurz, K. D., Main, B. W., Sandusky, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+model+of+arterial+thrombosis+induced+by+ferric+chloride.">Rat model of arterial thrombosis induced by ferric chloride.</a> Thromb Res. 60, 269-280 (1990).</li><li>Konstantinides, S., Schafer, K., Thinnes, T., Loskutoff, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasminogen+activator+inhibitor-1+and+its+cofactor+vitronectin+stabilize+arterial+thrombi+after+vascular+injury+in+mice.">Plasminogen activator inhibitor-1 and its cofactor vitronectin stabilize arterial thrombi after vascular injury in mice.</a> Circulation. 103, 576-583 (2001).</li><li>Leadley, R. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacodynamic+activity+and+antithrombotic+efficacy+of+RPR120844,+a+novel+inhibitor+of+coagulation+factor+Xa.">Pharmacodynamic activity and antithrombotic efficacy of RPR120844, a novel inhibitor of coagulation factor Xa.</a> J Cardiovasc Pharmacol. 34, 791-799 (1999).</li><li>Marsh Lyle, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+thrombin+inhibitor+efficacy+in+a+novel+rabbit+model+of+simultaneous+arterial+and+venous+thrombosis.">Assessment of thrombin inhibitor efficacy in a novel rabbit model of simultaneous arterial and venous thrombosis.</a> Thromb Haemost. 79, 656-662 (1998).</li><li>Farrehi, P. M., Ozaki, C. K., Carmeliet, P., Fay, W. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+arterial+thrombolysis+by+plasminogen+activator+inhibitor-1+in+mice.">Regulation of arterial thrombolysis by plasminogen activator inhibitor-1 in mice.</a> Circulation. 97, 1002-1008 (1998).</li><li>Robertson, J. O., Li, W., Silverstein, R. L., Topol, E. J., Smith, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deficiency+of+LRP8+in+mice+is+associated+with+altered+platelet+function+and+prolonged+time+for+in+vivo+thrombosis.">Deficiency of LRP8 in mice is associated with altered platelet function and prolonged time for in vivo thrombosis.</a> Thromb Res. 123, 644-652 (2009).</li><li>Gupta, N., Li, W., Willard, B., Silverstein, R. L., McIntyre, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proteasome+proteolysis+supports+stimulated+platelet+function+and+thrombosis.">Proteasome proteolysis supports stimulated platelet function and thrombosis.</a> Arterioscler Thromb Vasc Biol. 34, 160-168 (2014).</li><li>Srikanthan, S., Li, W., Silverstein, R. L., McIntyre, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exosome+poly-ubiquitin+inhibits+platelet+activation,+downregulates+CD36+and+inhibits+pro-atherothombotic+cellular+functions.">Exosome poly-ubiquitin inhibits platelet activation, downregulates CD36 and inhibits pro-atherothombotic cellular functions.</a> J Thromb Haemost. 12, 1906-1917 (2014).</li><li>Li, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thymidine+phosphorylase+participates+in+platelet+signaling+and+promotes+thrombosis.">Thymidine phosphorylase participates in platelet signaling and promotes thrombosis.</a> Circ Res. 115, 997-1006 (2014).</li><li>Le Menn, R., Bara, L., Samama, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrastructure+of+a+model+of+thrombogenesis+induced+by+mechanical+injury.">Ultrastructure of a model of thrombogenesis induced by mechanical injury.</a> J Submicrosc Cytol. 13, 537-549 (1981).</li><li>Li, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CD36+participates+in+a+signaling+pathway+that+regulates+ROS+formation+in+murine+VSMCs.">CD36 participates in a signaling pathway that regulates ROS formation in murine VSMCs.</a> J Clin Invest. 120, 3996-4006 (2010).</li><li>Re-examining Acute Eligibility for Thrombolysis Task Force. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review,+historical+context,+and+clarifications+of+the+NINDS+rt-PA+stroke+trials+exclusion+criteria:+Part+1:+rapidly+improving+stroke+symptoms.">Review, historical context, and clarifications of the NINDS rt-PA stroke trials exclusion criteria: Part 1: rapidly improving stroke symptoms.</a> Stroke. 44, 2500-2505 (2013).</li><li>Mumaw, M. M., de la Fuente, M., Arachiche, A., Wahl, J. K., Nieman, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+characterization+of+monoclonal+antibodies+against+Protease+Activated+Receptor+4+(PAR4).">Development and characterization of monoclonal antibodies against Protease Activated Receptor 4 (PAR4).</a> Thromb Res. 135, 1165-1171 (2015).</li><li>Mumaw, M. M., de la Fuente, M., Noble, D. N., Nieman, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+the+anionic+region+of+human+protease-activated+receptor+4+inhibits+platelet+aggregation+and+thrombosis+without+interfering+with+hemostasis.">Targeting the anionic region of human protease-activated receptor 4 inhibits platelet aggregation and thrombosis without interfering with hemostasis.</a> J Thromb Haemost. 12, 1331-1341 (2014).</li><li>Modery-Pawlowski, C. L., Kuo, H. H., Baldwin, W. M., Sen Gupta, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+platelet-inspired+paradigm+for+nanomedicine+targeted+to+multiple+diseases.">A platelet-inspired paradigm for nanomedicine targeted to multiple diseases.</a> Nanomedicine (Lond). 8, 1709-1727 (2013).</li><li>Anselmo, A. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Platelet-like+nanoparticles:+mimicking+shape,+flexibility,+and+surface+biology+of+platelets+to+target+vascular+injuries.">Platelet-like nanoparticles: mimicking shape, flexibility, and surface biology of platelets to target vascular injuries.</a> ACS Nano. 8, 11243-11253 (2014).</li><li>Modery, C. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heteromultivalent+liposomal+nanoconstructs+for+enhanced+targeting+and+shear-stable+binding+to+active+platelets+for+site-selective+vascular+drug+delivery.">Heteromultivalent liposomal nanoconstructs for enhanced targeting and shear-stable binding to active platelets for site-selective vascular drug delivery.</a> Biomaterials. 32, 9504-9514 (2011).</li><li>Woollard, K. J., Sturgeon, S., Chin-Dusting, J. P., Salem, H. H., Jackson, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Erythrocyte+hemolysis+and+hemoglobin+oxidation+promote+ferric+chloride-induced+vascular+injury.">Erythrocyte hemolysis and hemoglobin oxidation promote ferric chloride-induced vascular injury.</a> J Biol Chem. 284, 13110-13118 (2009).</li><li>Ciciliano, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resolving+the+multifaceted+mechanisms+of+the+ferric+chloride+thrombosis+model+using+an+interdisciplinary+microfluidic+approach.">Resolving the multifaceted mechanisms of the ferric chloride thrombosis model using an interdisciplinary microfluidic approach.</a> Blood. 126, 817-824 (2015).</li><li>Barr, J. D., Chauhan, A. K., Schaeffer, G. V., Hansen, J. K., Motto, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Red+blood+cells+mediate+the+onset+of+thrombosis+in+the+ferric+chloride+murine+model.">Red blood cells mediate the onset of thrombosis in the ferric chloride murine model.</a> Blood. 121, 3733-3741 (2013).</li><li>Dunne, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cadherin+6+has+a+functional+role+in+platelet+aggregation+and+thrombus+formation.">Cadherin 6 has a functional role in platelet aggregation and thrombus formation.</a> Arterioscler Thromb Vasc Biol. 32, 1724-1731 (2012).</li><li>Lockyer, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=GPVI-deficient+mice+lack+collagen+responses+and+are+protected+against+experimentally+induced+pulmonary+thromboembolism.">GPVI-deficient mice lack collagen responses and are protected against experimentally induced pulmonary thromboembolism.</a> Thromb Res. 118, 371-380 (2006).</li><li>Zhou, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+C-terminal+CGHC+motif+of+protein+disulfide+isomerase+supports+thrombosis.">The C-terminal CGHC motif of protein disulfide isomerase supports thrombosis.</a> J Clin Invest. , (2015).</li><li>Eckly, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+underlying+FeCl3-induced+arterial+thrombosis.">Mechanisms underlying FeCl3-induced arterial thrombosis.</a> J Thromb Haemost. 9, 779-789 (2011).</li><li>Day, S. M., Reeve, J. L., Myers, D. D., Fay, W. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+thrombosis+models.">Murine thrombosis models.</a> Thromb Haemost. 92, 486-494 (2004).</li><li>Cooley, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+models+of+thrombosis.">Murine models of thrombosis.</a> Thromb Res. 129 Suppl 2, S62-S64 (2012).</li><li>Gupta, N., Li, W., McIntyre, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deubiquitinases+Modulate+Platelet+Proteome+Ubiquitination,+Aggregation,+and+Thrombosis.">Deubiquitinases Modulate Platelet Proteome Ubiquitination, Aggregation, and Thrombosis.</a> Arterioscler Thromb Vasc Biol. 35, 2657-2666 (2015).</li><li>Konstantinides, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+antithrombotic+consequences+of+platelet+glycoprotein+Ibalpha+and+VI+deficiency+in+a+mouse+model+of+arterial+thrombosis.">Distinct antithrombotic consequences of platelet glycoprotein Ibalpha and VI deficiency in a mouse model of arterial thrombosis.</a> J Thromb Haemost. 4, 2014-2021 (2006).</li><li>Versteeg, H. H., Heemskerk, J. W., Levi, M., Reitsma, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+fundamentals+in+hemostasis.">New fundamentals in hemostasis.</a> Physiol Rev. 93, 327-358 (2013).</li><li>Yan, S. F., Mackman, N., Kisiel, W., Stern, D. M., Pinsky, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypoxia/Hypoxemia-Induced+activation+of+the+procoagulant+pathways+and+the+pathogenesis+of+ischemia-associated+thrombosis.">Hypoxia/Hypoxemia-Induced activation of the procoagulant pathways and the pathogenesis of ischemia-associated thrombosis.</a> Arterioscler Thromb Vasc Biol. 19, 2029-2035 (1999).</li><li>Rahaman, S. O., Li, W., Silverstein, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vav+Guanine+nucleotide+exchange+factors+regulate+atherosclerotic+lesion+development+in+mice.">Vav Guanine nucleotide exchange factors regulate atherosclerotic lesion development in mice.</a> Arterioscler Thromb Vasc Biol. 33, 2053-2057 (2013).</li><li>Silverstein, R. L., Li, W., Park, Y. M., Rahaman, S. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+cell+signaling+by+the+scavenger+receptor+CD36:+implications+in+atherosclerosis+and+thrombosis.">Mechanisms of cell signaling by the scavenger receptor CD36: implications in atherosclerosis and thrombosis.</a> Trans Am Clin Climatol Assoc. 121, 206-220 (2010).</li><li>Liu, J., Li, W., Chen, R., McIntyre, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circulating+biologically+active+oxidized+phospholipids+show+on-going+and+increased+oxidative+stress+in+older+male+mice.">Circulating biologically active oxidized phospholipids show on-going and increased oxidative stress in older male mice.</a> Redox Biol. 1, 110-114 (2013).</li></ol>

Den FeCl3-inducerad modell är en av de mest använda trombos modeller, som inte bara kan ge värdefull information om genetiska modifieringar på trombocytfunktion och trombos 7,8,16,19,31-33, men kan också vara ett värdefullt verktyg för utvärdering av terapeutiska föreningar och strategier för behandling och prevention av aterotrombotiska sjukdomar 11,17,34-37. Här har vi visat våra ändringar och förbättringar av denna modell och visade ytterligare bevis för nyttan av denn...

Arteriell trombos (blodpropp) är en vanlig komplikation vid många systemiska sjukdomar associerade med kronisk inflammation, inklusive ateroskleros, diabetes, fetma, cancer och kroniska autoimmuna reumatologiska sjukdomar. Tromber som förekommer i den arteriella cirkulationen stammen från olämpligt trombocytaktivering, aggregering och efterföljande koagulativa mekanismer, och är inblandade i hjärtinfarkt, stroke och extremiteter förlust. Kärlväggen är ett komplext system som omfattar flera celltyper och påverkas av en mängd yttre faktorer inklusive skjuvspänning, cirkulerande blodceller, hormoner och cytokiner, såväl som uttryck av antioxidant proteiner i kärlväggen. In vitro-försök kan inte replikera denna komplexa miljö och därför in vivo-studier med hjälp av djurmodeller är avgörande för att möjliggöra bättre förståelse för de mekanismer som är involverade i trombotiska störningar. Möss har visats ha simnande mekanismer för människor i form av trombos, åderförkalkning, inflammation och diabetes 1,2. Vidare kan transgena och knockoutmöss att skapas för att testa funktionen av specifika genprodukter i en komplex fysiologisk eller patologisk miljö. Sådana studier efterlikna mänsklig patologi och kan ge viktig mekanistisk information relaterad till upptäckten av nya vägar och terapier, samt ge viktiga detaljer i karakterisera läkemedelseffekter på trombos. Patologiska arteriella tromber uppstå på grund av endotelskiktet skada eller dysfunktion och exponering av blodströmmen till den subendoteliala matrisen 3,4. Olika trombos modeller har utvecklats för att inducera denna endotelskada såsom mekanisk skada, fotoreaktiva föreningen Rose Bengal-baserade oxidativ skada och laser skada 5. I detta spektrum, Järnklorid (FeCls 3) -inducerad kärlskada är en allmänt använd modell av trombos. Detta reagens närappliceras på den yttre aspekten av fartyg inducerar oxidativ skada på vaskulära celler 6-8, med förlust av endotel cellskyddet från cirkulerande blodplättar och komponenter i koagulationskaskaden. Den FeCl3 Modellen är enkel och känslig för både antikoagulerande och anti-trombocyter droger, och har utförts på halspulsådern och lårbensartärerna, hals vener och mesenteriska och cremasteric arterioler och venoler hos möss, råttor, marsvin och kaniner 6-15. En mätbar parameter i denna modell är den tid som förflutit från skada för att slutföra kärlocklusion, mätt som blodflödet upphör med en Doppler flödesmätare eller under direkt observation med intravital mikroskopi 6,7,9. Ett intervall av tider mellan 5 till 30 min har rapporterats i olika studier i C57BL6-möss 7-10,16, vilket antyder att FeCl 3 koncentrationer, typer av anestesi, kirurgiska tekniker, mus ålder, genomisk bakgrund, metoden för mätning av bLood flöde, och andra miljövariabler har betydande effekter i den här modellen. Denna breda variationen gör det svårt att jämföra studier från olika forskargrupper och kan göra detektion av subtila skillnader svårt. Med en vision att minimera sådana variabilitet och etablera ett likformigt reproducerbar in vivo modellsystem har vi förfinat FeCl3-inducerad halspulsådern modell som gör data mycket reproducerbara med minimal variation 6-10,16-19. I detta dokument beskriver vi och dela kunskaper och rapportera flera representativa experimentella exempel som kan dra nytta av denna modell.

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			<td height="40" style="height:40px;width:351px;">Metzenbaum Fino Scissors - Tungsten Carbide/Curved/Blunt-Blunt/14.5cm</td>
			<td style="width:159px;">Fine Science Tools&#160;</td>
			<td style="width:119px;">14519-14&#160;&#160;</td>
			<td>to dissect and separate soft tissue</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:351px;">Ultra Fine Hemostat - Smooth/Curved/12.5cm</td>
			<td style="width:159px;">Fine Science Tools&#160;</td>
			<td>13021-12</td>
			<td>to dissect and separate soft tissue</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Graefe Forceps - Serrated/Straight/10cm</td>
			<td style="width:159px;">Fine Science Tools&#160;</td>
			<td style="width:119px;">11050-10</td>
			<td>to dissect and separate soft tissue</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:351px;">Dumont #5 Fine Forceps - Biology Tips/Straight/Inox/11cm</td>
			<td style="width:159px;">Fine Science Tools&#160;</td>
			<td>11254-20&#160;</td>
			<td>Isolate vessel from surounding tissue</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:351px;">Dumont #5XL Forceps - Standard Tips/Straight/Inox/15cm</td>
			<td style="width:159px;">Fine Science Tools&#160;</td>
			<td>11253-10</td>
			<td>Isolate vessel from surounding tissue</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:351px;">Blunt Hook- 12cm/0.3mm Tip Diameter</td>
			<td style="width:159px;">Fine Science Tools&#160;</td>
			<td>10062-12</td>
			<td>Isolate vessel from surounding tissue</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:351px;">Castroviejo Micro Needle Holders</td>
			<td style="width:159px;">Fine Science Tools&#160;</td>
			<td>12061-02</td>
			<td>Needle holders</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:351px;">Suture Thread 4-0</td>
			<td style="width:159px;">Fine Science Tools&#160;</td>
			<td>18020-40</td>
			<td>For fix the incisors to the plate</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:351px;">Suture Thread 6-0</td>
			<td style="width:159px;">Fine Science Tools&#160;</td>
			<td>18020-60</td>
			<td>For all surgery and ligation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:351px;">Kalt Suture Needles</td>
			<td style="width:159px;">Fine Science Tools&#160;</td>
			<td>12050-03</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">rhodamine 6G&#160;</td>
			<td style="width:159px;">Sigma</td>
			<td>83697-1G</td>
			<td>To lebel platelets</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">FeCl3 (Anhydrous)</td>
			<td style="width:159px;">Sigma</td>
			<td style="width:119px;">12321</td>
			<td>To induce vessel injury</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Papaverine hydrochloride</td>
			<td style="width:159px;">Sigma</td>
			<td>P3510</td>
			<td>To inhibit gut peristalsis.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:351px;">Medline Surgical Instrument Sterilization Steam Autoclave Tapes</td>
			<td style="width:159px;">Medline</td>
			<td>111625</td>
			<td>To fix the mouse to the plate</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:351px;">Fisherbrand&#8482;&#160;Syringe Filters - Sterile 0.22&#181;m</td>
			<td style="width:159px;">Fisher</td>
			<td>09-720-004</td>
			<td>For sterlization of solutions injected to mice</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:351px;">Fisherbrand&#8482;&#160;Syringe Filters - Sterile 0.45&#181;m</td>
			<td style="width:159px;">Fisher</td>
			<td>09-719D</td>
			<td>To filter the FeCl3 solution</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:351px;">Sterile Alcohol Prep Pad</td>
			<td style="width:159px;">Fisher</td>
			<td>06-669-62</td>
			<td>To sterilize the surgical site</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:351px;">Agarose&#160;</td>
			<td style="width:159px;">BioExpress</td>
			<td>E-3120-500</td>
			<td>To make gel stage</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:351px;">Leica DMLFS fluorescent microscope</td>
			<td style="width:159px;">Leica</td>
			<td style="width:119px;"></td>
			<td>Intravital microscope</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:351px;">GIBRALTAR Platform and X-Y Stage System</td>
			<td style="width:159px;">npi electronic GmbH</td>
			<td style="width:119px;"></td>
			<td style="width:325px;">http://www.npielectronic.de/products/micropositioners/burleigh/gibraltar.html</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:351px;">Streampix version 3.17.2 software</td>
			<td style="width:159px;">NorPix</td>
			<td style="width:119px;"></td>
			<td>https://www.norpix.com/</td>
		</tr>
	</tbody>
</table>

ferric chloride-induced murine thrombosis models

Arterial thrombosis (blood clot) is a common complication of many systemic diseases associated with chronic inflammation, including atherosclerosis, diabetes, obesity, cancer and chronic autoimmune rheumatologic disorders. Thrombi are the cause of most heart attacks, strokes and extremity loss, making thrombosis an extremely important public health problem. Since these thrombi stem from inappropriate platelet activation and subsequent coagulation, targeting these systems therapeutically has important clinical significance for developing safer treatments. Due to the complexities of the hemostatic system, in vitro experiments cannot replicate the blood-to-vessel wall interactions; therefore, in vivo studies are critical to understand pathological mechanisms of thrombus formation. To this end, various thrombosis models have been developed in mice. Among them, ferric chloride (FeCl3) induced vascular injury is a widely used model of occlusive thrombosis that reports platelet activation and aggregation in the context of an aseptic closed vascular system. This model is based on redox-induced endothelial cell injury, which is simple and sensitive to both anticoagulant and anti-platelets drugs. The time required for the development of a thrombus that occludes blood flow gives a quantitative measure of vascular injury, platelet activation and aggregation that is relevant to thrombotic diseases. We have significantly refined this FeCl3-induced vascular thrombosis model, which makes the data highly reproducible with minimal variation. Here we describe the model and present representative data from several experimental set-ups that demonstrate the utility of this model in thrombosis research.

Halspulsådern trombosmodell Hos möss med C57BL6 bakgrund rekommenderar vi att du använder 7,5% FeCl3 att behandla fartyget under 1 minut som utgångspunkt. Under behandling av 7,5% FeCl3, är gränserna för skadade området och normal kärlväggen lätt identifieras under mikroskop (se online video 1), vilket tyder på att endotelskiktet skadades betydligt. Tromb bildades omedelbart vid FeCl3 behandling, och ob...

Watch this Scientific Journal Video about Ferric Chloride-induced Murine Thrombosis Models at JoVE.com

Ferric Chloride-induced Murine Thrombosis Models

Vi rapporterar en raffinerad förfarandet för järnklorid (FeCl3) -inducerade modeller trombos på hals och mesenterica liksom ven, kännetecknad effektivt med intravital mikroskopi för att övervaka tid att ocklusiv trombbildning.

Järnklorid-inducerad Murine Trombos modeller

Arterial thrombosis (blood clot) is a common complication of many systemic diseases associated with chronic inflammation, including atherosclerosis, ...

Research

JoVE Journal

Medicine

Electrolytic Inferior Vena Cava Model (EIM) of Venous Thrombosis

Electrolytic Inferior Vena Cava Model EIM of Venous Thrombosis

Elektrolytisk Inferior Vena Cava Model (EIM) för venös trombos

Animal models serve a vital role in deep venous thrombosis (DVT) research in order to study thrombus formation, thrombus resolution and to test potential ...

Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

Mus Komplett Stasis modell av sämre Vena Cava Trombos

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE).  In the United States (U.S.), the high morbidity and ...

Segmentation and Measurement of Fat Volumes in Murine Obesity Models Using X-ray Computed Tomography

Segmentering och Mätning av FAT-volymer i murina fetma modeller med hjälp av X-ray datortomografi

Obesity is associated with increased morbidity and mortality as well as reduced metrics in quality of life.1 Both environmental and genetic factors are ...

Quantitative Analysis and Characterization of Atherosclerotic Lesions in the Murine Aortic Sinus

Kvantitativ Analys och karakterisering av aterosklerotiska lesioner i murina Aorta Sinus

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have ...

Intrastriatal Injection of Autologous Blood or Clostridial Collagenase as Murine Models of Intracerebral Hemorrhage

Intrastriatal Injektion av autologt blod eller Clostridial kollagenas som musmodeller av intracerebral blödning

Intracerebral hemorrhage (ICH) is a common form of cerebrovascular disease and is associated with significant morbidity and mortality. Lack of effective ...

Orthotopic Injection of Breast Cancer Cells into the Mammary Fat Pad of Mice to Study Tumor Growth.

Orthotopic Injektion av bröstcancerceller i juverfett Pad av möss att studera tumörtillväxt.

Breast cancer growth can be studied in mice using a plethora of models. Genetic manipulation of breast cancer cells may provide insights into the ...

Adapting Human Videofluoroscopic Swallow Study Methods to Detect and Characterize Dysphagia in Murine Disease Models

Anpassning Human Videofluoroscopic Swallow studiemetoder för att upptäcka och karakterisera Dysfagi i murina Sjukdom modeller

This study adapted human videofluoroscopic swallowing study (VFSS) methods for use with murine disease models for the purpose of facilitating ...

High-frequency Ultrasound Imaging of Mouse Cervical Lymph Nodes

Högfrekvent ultraljud avbildning av Mouse Cervical lymfkörtlar

High-frequency ultrasound (HFUS) is widely employed as a non-invasive method for imaging internal anatomic structures in experimental small animal ...

A Multicenter MRI Protocol for the Evaluation and Quantification of Deep Vein Thrombosis

En Multi MRI protokoll för utvärdering och kvantifiering av djup ventrombos

We evaluated a magnetic resonance venography (MRV) approach with gadofosveset to quantify total thrombus volume changes as the principal criterion for ...

Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

Järnklorid-inducerad Trombos musmodell på Carotid Artery och tarmkäx Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...