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.

所有的过程和动物的操作都按照美国公共卫生服务政策上的人文关怀和动物的使用 ，以及NIH 指南照顾和批准了克利夫兰诊所的机构动物护理和使用委员会（IACUC） 实验动物使用 。 1.准备： <ol><li> 荧光染料用于标记血小板 <ol><li>制备罗丹明6G溶液，0.5毫克/毫升，在盐水和消毒用0.22微米的过滤器将溶液。 </li></ol></li><li> 三氯化铁3溶液 <o...

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所述的FeCl 3诱导的模型是最广泛使用的血栓形成的模型，它不仅可以提供有关对血小板功能和血栓形成7,8,16,19,31-33遗传修饰的有价值的信息中的一个，但也可以是一个有价值的工具用于治疗化合物和战略动脉粥样硬化性疾病11,17,34-37的治疗和预防的评价。在这里，我们已经表明我们的修改和该模型的改进和表明该技术中，这是足够敏感以确定抗凝物（tPA），抗血小板药物?...

动脉血栓形成（血块）是与慢性炎症，包括动脉粥样硬化，糖尿病，肥胖症，癌症和慢性自身免疫性风湿病相关的许多全身性疾病的常见并发症。发生在不适当的血小板活化，聚集和后续机制混凝动脉循环茎，血栓有牵连的心脏发作，中风和肢体损失。血管壁是一个复杂的系统，它包括多种细胞类型和是由外在因素，包括剪切应力多种影响，循环血细胞，激素和细胞因子，以及抗氧化蛋白表达在血管壁。 体外实验不能复制这种复杂的环境，因此， 在使用动物模型的体内研究是至关重要的，以允许更好地理解涉及血栓性疾病的机制。 小鼠已经显示出具有卡ILAR机制对人类中的血栓形成，动脉粥样硬化，炎症和糖尿病1,2-条款。此外，可以创建转基因和基因敲除小鼠测试的特定基因产物的功能在一个复杂的生理或病理环境。这样的研究模仿人类病理学，并且可以提供有关的新途径和疗法，发现重要的机械信息以及在上血栓形成表征药物作用提供重要的信息。 病理动脉血栓的发生是由于内皮层损伤或功能障碍和血液流暴露于内皮下矩阵3,4。各种血栓形成模型已经发展到诱导这种内皮损伤，如机械性损伤，基于孟加拉-光反应性化合物玫瑰氧化损伤和激光损伤5。在这个频谱，氯化铁（ 氯化铁）诱导的血管损伤是血栓形成的一个广泛使用的模型。该试剂时，施加到容器的外表方面诱导对血管细胞6-8氧化损伤，与来自循环血小板和凝血级联组分内皮细胞保护的丧失。所述的FeCl 3模型简单到两个抗凝和抗血小板药物敏感，并已在颈动脉和股动脉，颈静脉和肠系膜和提睾动脉和小静脉在小鼠，大鼠，豚鼠和兔6-15进行。 在此模型中的一个可测量参数是从损伤的经过时间来完成血管闭塞，作为血流停止用多普勒血流计或根 ​​据与活体显微镜6,7,9直接观察来测定。的5至30分钟之间倍一系列已报道在C57BL6小鼠7-10,16不同的研究，表明的FeCl 3浓度，种类麻醉，手术技术，小鼠年龄，基因组背景，测量的B法的lood流，和其他环境变量具有在该模型显著影响。这个很大的可变性使得难以研究从不同的研究小组比较，并且可以使细微的差别检测变得困难。 具有远见尽量减少这种变异，并建立一个在体内模型系统统一重复性好，我们已经完善了氯化铁诱发颈动脉模型，从而以最小的变化6-10,16-19高重现性的数据。在本文中，我们描述和分享技能和报告，可以从这个模式中受益，几个有代表性的实验例。

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		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="20">
			<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>
		</tr>
		<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>
		</tr>
		<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.

颈动脉血栓形成模型 在C57BL6背景小鼠中，我们建议使用7.5％的氯化铁治疗血管1分钟为出发点。下的FeCl 3处理的7.5％，受伤的区域和正常血管壁的边界显微镜下很容易识别（参见在线视频1），这表明内皮层显著损坏。血栓形成，立即经氯化铁处理，损伤后的所有WT C57BL6小鼠1分钟观察。最初形成的血栓是不稳定的，它?...

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

Ferric Chloride-induced Murine Thrombosis Models

我们报告的氯化铁（氯化铁3）颈动脉和肠系膜动脉血栓诱发款精致的程序以及静脉，其特征有效地利用活体显微镜监测的时间来闭塞血栓的形成。

三氯化铁诱导小鼠模型的血栓形成

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

电解下腔静脉静脉血栓形成的模型（EIM）

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

科研

医学

Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

鼠标完成血瘀模型的下腔静脉血栓

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

利用X射线计算机断层扫描在小鼠肥胖模型的FAT卷的分割和测量

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

自体血液或梭菌胶原酶脑内注射为脑出血的小鼠模型

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.

注射原位乳腺癌细胞到小鼠乳腺脂肪垫来研究肿瘤的生长。

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

适应人类Videofluoroscopic燕子研究的方法来检测和检定吞咽困难的小鼠疾病模型

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

鼠标颈淋巴结高频超声成像

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

深静脉血栓形成的评价和定量的多中心MRI协议

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

三氯化铁诱发血栓形成小鼠模型上的颈动脉和肠系膜血管

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