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.

Alle prosedyrer og manipulasjoner av dyr har blitt godkjent av Institutional Animal Care og Bruk komiteer (IACUC) av The Cleveland Clinic i samsvar med USA Public Health Service politikk på Humane Stell og bruk av dyr, og NIH Guide for Pleie og bruk av forsøksdyr. 1. Forberedelser: <ol><li> Fluorescerende fargestoff for merking Blodplater <ol><li> Fremstille rodamin 6G oppløsning, 0,5 mg / ml i saltvann og sterilisere løsningen med 0,22 um filter. </li></ol></li><li> <str...

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Det FeCl3-indusert modell er en av de mest brukte tromboser modeller, som ikke bare kan gi verdifull informasjon om genetiske modifikasjoner på blodplatefunksjon og trombose 7,8,16,19,31-33, men kan også være et verdifullt verktøy for evaluering av terapeutiske forbindelser og strategier for behandling og forebygging av sykdommer aterotrombotiske 11,17,34-37. Her har vi vist våre modifikasjoner og forbedringer av denne modell og viste ytterligere bevis på anvendeligheten av denne t...

Arteriell trombose (blodpropp) er en vanlig komplikasjon ved mange systemiske sykdommer assosiert med kronisk betennelse, inkludert aterosklerose, diabetes, fedme, kreft og kroniske autoimmune revmatologiske lidelser. Tromber som oppstår i den arterielle sirkulasjonen stammer fra upassende plateaktivering, aggregering og påfølgende coagulatory mekanismer, og er innblandet i hjerteinfarkt, slag og ekstremiteten tap. Karveggen er et komplekst system som inneholder flere celletyper og er påvirket av en rekke av ytre faktorer, inkludert skjærspenning, sirkulerende blodceller, hormoner og cytokiner, så vel som ekspresjon av antioksidant proteiner i karveggen. In vitro forsøk kan ikke replikere dette komplekse miljø og derfor in vivo-studier som anvender dyremodeller er kritiske for å gi bedre forståelse av mekanismene som er involvert i trombotiske lidelser. Mus har vist seg å ha simILAR mekanismer til mennesker i form av trombose, aterosklerose, betennelser og diabetes 1,2. Videre kan transgene mus og knockout opprettes for å teste funksjonen av spesifikke genprodukter i et komplekst fysiologisk eller patologisk miljø. Slike studier etterligne menneskelig patologi og kan gi viktig mekanistisk informasjon knyttet til oppdagelsen av nye stier og terapi, samt gi viktige opplysninger for å karakterisere virkninger av rusmidler på trombose. Patologiske arterielle tromber oppstå på grunn til endotele lag skade eller dysfunksjon og eksponering av blodstrømmen til subendotel-matriksen 3,4. Ulike trombose modeller har blitt utviklet for å stimulere denne endothelial skader som mekanisk skade, fotoreaktive forbindelse Rose Bengal-baserte oksidativ skade og laser skade fem. I dette spektrum, Ferroklorid (FeCl3) indusert vaskulær skade er en mye brukt modell av trombose. Denne reagensen nårpåført den ytre del av fartøyer induserer oksidativ skade på vaskulære celler 6-8, med tap av endotel celle beskyttelse fra sirkulerende blodplater og komponenter i koagulasjonskaskaden. Det FeCl3-modellen er enkel og følsomme for både antikoagulant og anti-blodplater stoffer, og har vært utført på arteria carotis og femorale arterier, vena jugularis, og mesenterisk og cremasteric arterioler og venuler i mus, rotter, marsvin og kaniner 6-15. En målbar parameter i denne modellen er den tiden som går fra skade til å fullføre fartøy okklusjon, målt som blodstrømmen opphør med en Doppler strømningsmåler eller under direkte observasjon med intramikros 6,7,9. En rekke ganger mellom 5 til 30 minutter er blitt rapportert i forskjellige studier på C57BL6 mus 7-10,16, noe som tyder på at FeCl3 konsentrasjoner typer anestesi, kirurgiske teknikker, mus alder, genomisk bakgrunn, fremgangsmåte for måling blood flyt, og andre miljøvariabler har betydelige effekter i denne modellen. Denne brede variasjonen gjør det vanskelig å sammenligne studier fra ulike fagmiljøer og kan gjøre deteksjon av små forskjeller vanskelig. Med en visjon om å minimere slike variabilities og etablere en enhetlig reproduserbar in vivo modell system, har vi videreutviklet FeCl3 indusert halspulsåren modell som gjør dataene svært reproduserbare med minimal variasjon 6-10,16-19. I denne artikkelen beskriver vi og dele kompetanse og rapportere flere representative eksperimentelle eksempler som kan dra nytte av denne modellen.

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			<td height="20" style="height:20px;width:351px;">Surgical Scissors - Tungsten Carbide</td>
			<td style="width:159px;">Fine Science Tools&#160;</td>
			<td style="width:119px;">14502-14</td>
			<td style="width:325px;">cut and hold skin</td>
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		<tr height="20">
			<td height="20" style="height:20px;width:351px;">Micro-Adson Forceps - Serrated/Straight/12cm</td>
			<td style="width:159px;">Fine Science Tools&#160;</td>
			<td>11018-12</td>
			<td>cut and hold skin</td>
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		<tr height="40">
			<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.

Carotis arteietrombose Model Hos mus med C57BL6 bakgrunn, anbefaler vi å bruke 7,5% FeCl3 å behandle fartøyet i 1 min som utgangspunkt. Under behandling av 7,5% FeCl3, er grensene til skadde området og normal åreveggen lett identifiseres i henhold mikroskop (Se online video 1), noe som tyder på at endothelial laget ble betydelig skadet. Trombi som dannes umiddelbart etter FeCl3 behandling, og blir observert i...

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

Ferric Chloride-induced Murine Thrombosis Models

Vi rapporterer en raffinert prosedyre av jernklorid (FeCl3) indusert trombose modeller på carotis og mesenteric arterie samt vene, preget effektivt ved hjelp av intravital mikroskopi for å overvåke tid til okklusiv tromber formasjon.

Jernklorid-indusert Murine Trombose 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

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

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 og Måling av fett Volumene i murine Fedme modeller med X-ray Datatomografiscanning

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

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 Injeksjon av Autolog blod eller clostridial Collage som murine modeller av intracerebral blødning

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 Injeksjon av brystkreft celler inn i Mammary Fat Pad av mus å studere tumorvekst.

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

Tilpasse menneske Videofluoroscopic Swallow Study Metoder for å påvise og karakterisere Dysfagi i murine sykdomsmodeller

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

Høyfrekvente ultralydavbildning av Muse Cervical lymfeknuter

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 for evaluering og Kvantifisering av dyp venetrombose

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

Jernklorid-indusert tromboser Mouse Model på arteria carotis og mesenterium Vessel

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