Name: ferric chloride-induced murine thrombosis models
Uploaded: 2016-09-05T13:00:00
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.

Tutte le procedure e le manipolazioni di animali sono stati approvati dalla Institutional Animal Care e utilizzo Comitati (IACUC) di The Cleveland Clinic in accordo con gli Stati Uniti la politica di servizio sanitario pubblico sulla cura umana e all&#39;uso degli animali, e la Guida NIH per la cura e L&#39;uso di animali da laboratorio. 1. Preparazione: <ol><li> Colorante fluorescente per l&#39;etichettatura Piastrine <ol><li> Preparare rodamina 6G soluzion...

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Il modello di FeCl 3 indotta è uno dei modelli più utilizzati trombosi, che possono non solo fornire preziose informazioni modificazioni genetiche sulla funzione piastrinica e la trombosi 7,8,16,19,31-33, ma può anche essere uno strumento prezioso per la valutazione di composti terapeutici e strategie per il trattamento e la prevenzione delle malattie aterotrombotici 11,17,34-37. Qui abbiamo dimostrato le nostre modifiche e perfezionamenti di questo modello e ha mostrato ulteriori pro...

trombosi arteriosa (coagulo di sangue) è una complicanza comune di molte malattie sistemiche associate a infiammazione cronica, tra cui l&#39;aterosclerosi, il diabete, l&#39;obesità, cancro e patologie reumatologiche autoimmuni croniche. Trombi che si verificano nel gambo circolazione arteriosa da piastrine di attivazione inadeguato, l&#39;aggregazione e successive meccanismi coagulatoria, e sono implicati in attacchi di cuore, ictus e perdita di estremità. La parete del vaso è un sistema complesso che comprende diversi tipi di cellule ed è influenzata da molteplici fattori estrinseci compreso sollecitazione di taglio, le cellule del sangue, ormoni e citochine, così come espressione di proteine ​​antiossidanti circolanti nella parete del vaso. In vitro esperimenti non possono replicarsi questo ambiente complesso e, pertanto, in studi in vivo su modelli animali sono fondamentali per consentire una migliore comprensione dei meccanismi coinvolti nei disordini trombotici. I topi hanno dimostrato di avere simmeccanismi ili per gli esseri umani in termini di trombosi, arteriosclerosi, infiammazione e il diabete 1,2. Inoltre, i topi transgenici e knockout possono essere creati per testare la funzione di prodotti genici specifici in un fisiologico complesso o un ambiente patologico. Tali studi imitano patologia umana e possono fornire importanti informazioni relative al meccanicistico scoperta di nuovi percorsi e terapie, oltre a fornire dettagli importanti nel caratterizzare gli effetti della droga sulla trombosi. Trombi arteriosi patologiche si verificano a causa di lesioni endoteliali strato o disfunzione e l&#39;esposizione del flusso sanguigno alla matrice sottoendoteliale 3,4. Vari modelli trombosi sono stati sviluppati per indurre il danno endoteliale, come lesioni meccaniche, danno ossidativo composto fotoreattivo Rosa Bengala-based e lesioni a laser 5. In questo spettro, cloruro ferrico (FeCl 3) indotta danno vascolare è un modello ampiamente utilizzato di trombosi. Questo reagente quandoapplicata all&#39;aspetto esterno delle navi induce danno ossidativo alle cellule vascolari 6-8, con perdita di protezione delle cellule endoteliali dalle piastrine e componenti della cascata della coagulazione circolante. Il modello di FeCl 3 è semplice e sensibile sia anticoagulanti e antipiastrinici farmaci, ed è stata eseguita su arterie carotidee e femorali, vene giugulari e mesenterici ed arteriole cremasterico e venule nei topi, ratti, cavie e conigli 6-15. Un parametro misurabile in questo modello è il tempo trascorso da un infortunio per completare l&#39;occlusione nave, misurata come la cessazione del flusso sanguigno con un misuratore di portata Doppler o sotto l&#39;osservazione diretta con intravitale 6,7,9 microscopia. Una gamma di volte tra i 5 ei 30 minuti è stato riportato in diversi studi in topi C57Bl6 7-10,16, suggerendo che FeCl 3 concentrazioni, tipi di anestesia, tecniche chirurgiche, l&#39;età del mouse, sfondo genomica, metodo di misurazione Bflusso lood, e di altre variabili ambientali hanno effetti significativi in ​​questo modello. Questa ampia variabilità rende difficile il confronto tra gli studi di diversi gruppi di ricerca e può rendere la rilevazione di sottili differenze difficile. Con una visione per ridurre al minimo tali variabilità e stabilire un modo uniforme e riproducibile nel sistema modello in vivo, abbiamo affinato il modello dell&#39;arteria carotide FeCl 3 indotta che rende i dati altamente riproducibili con minime variazioni 6-10,16-19. In questo lavoro descriviamo e condividere le competenze e riportiamo alcuni esempi rappresentativi sperimentali che possono beneficiare di questo modello.

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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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			<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.

Carotide Trombosi Modello Nei topi con C57BL6 sfondo, si consiglia di utilizzare 7,5% FeCl 3 per trattare il vaso per 1 min come punto di partenza. Sotto trattamento del 7,5% FeCl 3, i confini della zona feriti e parete dei vasi normali sono facilmente identificabili al microscopio (vedi video in linea 1), suggerendo che lo strato endoteliale è stata significativamente danneggiata. Il trombi formata subito dopo FeCl 3</...

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

Ferric Chloride-induced Murine Thrombosis Models

Riportiamo una procedura raffinata del cloruro ferrico (FeCl 3) modelli trombosi -indotta sulla carotide e dell&#39;arteria mesenterica così come la vena, caratterizzato in modo efficiente usando la microscopia intravitale per monitorare il tempo di occlusiva la formazione di trombi.

Ferrico I modelli murini Trombosi indotta da cloruri

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

Elettrolitico vena cava inferiore Model (EIM) della Trombosi Venosa

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

Ricerca

Medicina

Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

Modello del mouse completa stasi della vena cava inferiore Trombosi

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

Segmentazione e misurazione dei volumi FAT nei modelli murini di obesità Utilizzo di X-ray Tomografia Computerizzata

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

Iniezione intrastriatali di sangue autologo o Clostridial Collagenase come modelli murini di emorragia intracerebrale

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.

Iniezione ortotopico di cellule di carcinoma mammario nel mammaria Fat Pad di Mice per studiare la crescita del tumore.

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

Adattamento umani videofluoroscopia Rondine metodi di studio per individuare e caratterizzare disfagia in modelli di malattia Murine

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

Ad alta frequenza di Ecografia del mouse cervicali Linfonodi

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

Multicentrico MRI protocollo per la valutazione e la quantificazione della trombosi venosa profonda

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

Cloruro ferrico-indotta Trombosi modello murino sulla carotide e Mesentere Vessel

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