Name: primary orthotopic glioma xenografts recapitulate infiltrative growth and isocitrate dehydrogenase i mutation
Uploaded: 2014-01-14T13:45:00
Description: Watch this Scientific Journal Video about Primary Orthotopic Glioma Xenografts Recapitulate Infiltrative Growth and Isocitrate Dehydrogenase I Mutation at JoVE.com

我们特别感谢范德比尔特大学医学中心的患者，他们为分子神经外科组织库提供了宝贵的研究材料。我们感谢那些建立和维护组织银行的人，里德·汤普森MD（首席调查员），樱桃金纳德RN（研究护士）和拉里A.皮尔斯MS（经理）。组织学服务部分由范德比尔特大学医学中心 （VUMC） 转化病理共享资源（由 5P30 CA068485 奖支持范德比尔特-英格拉姆癌症中心）执行。这项工作得到了国家发展局（1R21NS070139）、伯劳斯威康基金和VMC发展基金向MKC提供的赠款的支持。MKC 由退伍军人事务部、退伍军人健康管理局、研究和发展办公室、生物医学实验室研究和发展部通过赠款 1 I01 BX000744-01 获得支持。内容不代表退伍军人事务部或美国政府的意见。

1. 肿瘤细胞悬架的准备
注： 需要为使用患者材料和动物提供适当的机构批准，以建立和维持原发性正畸胶质瘤异种移植物。在范德比尔特大学医学中心，在患者同意下收集超过诊断所需量的肿瘤材料，用于研究组织库。标本标本标注带有随机的 5 位 REDcap 数据库编号，并删除所有患者特定标识符。REDcap 数据库包含组织存储库中每个标本的分离临床数据，包括性别...

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培养细胞系、异种移植和转基因小鼠是模拟胶质瘤的最常见方法，每个模型系统3，13，14都有明显的益处和局限性。原位正交胶质瘤异种移植的相关好处包括渗透性生长，这些生长是弥漫性胶质瘤的典型代表，以及保留遗传改变和重要的信号机制，这些机制在培养的胶质瘤细胞中可能极其难以维持。例如，isocitrate脱氢酶突变和声波刺猪配体生产可以维持在原位矫形异种格拉夫特...

恶性胶质瘤是发生在大脑和脊髓的中枢神经系统的原发性胶质肿瘤。胶质瘤由世界卫生组织（世卫组织）根据组织学上与星形细胞、寡头细胞或子细胞的相似性进行分类，然后根据恶性病理特征进行数值分级（I至IV）。最常见的组织学亚型是天体细胞瘤、寡头性腺胶质瘤和混合寡头肌细胞瘤。包括世卫组织二至四级在内的恶性胶质瘤的特点是侵入性生长和对现有疗法的顽固不化。在美国，每年约有15，750人被诊断为恶性胶质瘤，估计有12，740名患者死于这种疾病。这些统计数字突出了恶性胶质瘤的特别致命性，以及提高疗效的重要需要。
癌症模型对于研究肿瘤生物学和疗法至关重要。人类癌细胞系是体外操纵和体外异种移植研究（继发性异种移植）1的重要第一步。然而，标准癌细胞培养经历表型和基因型转换2-4，可能无法恢复在继发性异种移植5。此外，基因改变，如异辛酸盐脱氢酶（IDH）突变6，独特的干细胞种群7和对关键信号通路8的依赖，可能会在癌细胞培养中消失。基因组特征可以在癌症领域培养中更好地保持，但仍不能完全反映原肿瘤的基因型2，3。直接正交移植否定了体外培养的影响，提供了适当的微环境，并保留了肿瘤启动细胞9，10的完整性。因此，原生异种移植物是一种强大而相关的临床前模型，用于严格测试靶向剂，以帮助合理设计未来的临床试验5，11，12。

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			Phosphate buffered saline
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			Life Technologies
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			14040-133
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			Papain dissociation system
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			Worthington Biochemical Corp.
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			LK003150
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			Trypan blue solution 0.4%
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			Life Technologies
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			15250061
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			Ketamine HCl
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			Obtained from institutional pharmacy or local veterinary supply company
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			Xylazine HCl
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			Ketoprofen
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			Ophthalmic ointment
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			Povidone-iodine
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			Fisher Scientific
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			190061617
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			Cryopreservation medium and proliferation supplement
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			StemCell Technologies
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			05751
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			0.2% Heparin sodium salt in PBS
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			StemCell Technologies
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			07980
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			Penicillin-streptomycin
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			<td style="width:142px;">
			Life Technologies
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			15140-122
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			Dimethyl sulfoxide
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			Sigma-Aldrich
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			D6250-5X10ML
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			NOD.Cg-Prkdcscid I/2rgtm1Wjl/SzJ mice
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			The Jackson Laboratory
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			005557
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			NSG mice
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			Anti-human vimentin antibody
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			Dako
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			M7020
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			Use 1:200 to 1:800
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			Anti-human IDH1 R132H antibody
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			<td style="width:142px;">
			Dianova
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			DIA-H09
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			Use 1:100 to 1:400
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			Comments
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			Centrifuge with swinging bucket rotor
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			Pipetter with dispensing speed control
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			Disposable hemocytometer
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			<td style="width:140px;">
			Fisher Scientific
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			<td style="width:143px;">
			22-600-100
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			Sterile surgical gloves
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			<td style="width:140px;">
			Fisher Scientific
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			<td style="width:143px;">
			11-388128
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			Disposable gown
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			Fisher Scientific
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			18-567
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			Surgical mask
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			Fisher Scientific
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			19-120-1256
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			Tuberculin syringe
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			BD
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			305620
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			Alcohol pads
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			Fisher Scientific
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			22-246-073
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			Portable electronic scale
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			Fisher Scientific
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			01-919-33
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			Zoom stereomicroscope
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			Surgical clipper
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			Stoelting
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			51465
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		<tr>
			<td style="width:210px;">
			Scalpel handle
			</td>
			<td style="width:140px;">
			Fine Science Tools
			</td>
			<td style="width:143px;">
			10003-12
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">
			Scalpel blades, #10
			</td>
			<td style="width:140px;">
			
			</td>
			<td style="width:143px;">
			
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">
			Stereotaxic instrument
			</td>
			<td style="width:140px;">
			Stoelting
			</td>
			<td style="width:143px;">
			51730
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">
			High-speed drill
			</td>
			<td style="width:140px;">
			Stoelting
			</td>
			<td style="width:143px;">
			51449
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">Drill bit, 0.6 mm</td>
			<td style="width:140px;">Stoelting</td>
			<td style="width:143px;">514552</td>
			<td style="width:97px;"></td>
		</tr>
		<tr>
			<td style="width:210px;">
			Hamilton syringe
			</td>
			<td style="width:140px;">
			Hamilton
			</td>
			<td style="width:143px;">
			80336
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">
			Autoclip, 9 mm
			</td>
			<td style="width:140px;">
			BD
			</td>
			<td style="width:143px;">
			427630
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">
			Circulating water warming pad
			</td>
			<td style="width:140px;">
			Kent Scientific
			</td>
			<td style="width:143px;">
			TP-700
			TP-1215EA
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">
			Hot bead dry sterilizer
			</td>
			<td style="width:140px;">
			Kent Scientific
			</td>
			<td style="width:143px;">
			INS300850
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">
			Surgical scissors
			</td>
			<td style="width:140px;">
			Fine Science Tools
			</td>
			<td style="width:143px;">
			14101-14
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">
			Fine scissors
			</td>
			<td style="width:140px;">
			Fine Science Tools
			</td>
			<td style="width:143px;">
			14094-11
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">
			Spring scissors
			</td>
			<td style="width:140px;">
			Fine Science Tools
			</td>
			<td style="width:143px;">
			15018-10
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">
			Dumont forceps
			</td>
			<td style="width:140px;">
			Fine Science Tools
			</td>
			<td style="width:143px;">
			11251-30
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">
			Semimicro spatulas
			</td>
			<td style="width:140px;">
			Fisher Scientific
			</td>
			<td style="width:143px;">
			14374
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">
			Mouse brain slicer matrix
			</td>
			<td style="width:140px;">
			Zivic Instruments
			</td>
			<td style="width:143px;">
			BSMAS002-1
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
		<tr>
			<td style="width:210px;">
			Cryogenic storage vials
			</td>
			<td style="width:140px;">
			Fisher Scientific
			</td>
			<td style="width:143px;">
			12-567-501
			</td>
			<td style="width:97px;">
			
			</td>
		</tr>
	</tbody>
</table>

primary orthotopic glioma xenografts recapitulate infiltrative growth and isocitrate dehydrogenase i mutation

恶性胶质瘤是一组具有明显临床和分子特征的高度渗透性胶质肿瘤的异质性群体。原位矫形异种格拉夫特在前科动物模型中重新概括恶性胶质瘤亚型的组织病理学和分子特征。为了模拟世卫组织在移植检测中的I级和IV级恶性胶质瘤，人类肿瘤细胞被异种移植到免疫功能低下小鼠的正交部位——大脑中。与利用培养肿瘤细胞的继发性异种移植物相比，人类胶质瘤细胞与被切除的标本分离，在组织培养中无需事先通过移植即可产生原发性异种移植。本报告中的程序详细说明了肿瘤样本准备、颅内移植到免疫功能低下的小鼠、肿瘤移植监测以及肿瘤收获，供随后进入受体动物或进行分析。肿瘤细胞准备需要2小时，外科手术需要20分钟/动物。

分离的胶质瘤细胞被直接移植到免疫功能低下的小鼠的大脑中，以获得原位正畸异种移植线。每个肿瘤标本在移植前被分配一个随机数字，作为去除受保护的健康信息的分离过程的一部分。为此，我们使用 5 位 REDcap 数据库编号。 图1 说明了从胶质母细胞瘤（GBM 17182）与同心酸脱氢酶1（IDH1）突变精氨酸132到组织丁（R132H）建立异种移植线的过程和命名。移植后，肿瘤识别号中添加"X"?...

Watch this Scientific Journal Video about Primary Orthotopic Glioma Xenografts Recapitulate Infiltrative Growth and Isocitrate Dehydrogenase I Mutation at JoVE.com

Primary Orthotopic Glioma Xenografts Recapitulate Infiltrative Growth and Isocitrate Dehydrogenase I Mutation

恶性胶质瘤是一组具有明显临床和分子特征的高度渗透性胶质肿瘤的异质性群体。原位矫形异种格拉夫特在前科动物模型中重新概括恶性胶质瘤亚型的组织病理学和分子特征。

原正畸胶质血异草回顾渗透生长和同源脱氢酶 I 突变

Malignant gliomas constitute a heterogeneous group of highly infiltrative glial neoplasms with distinct clinical and molecular features. Primary ...

Research

JoVE Journal

Medicine

An Orthotopic Model of Murine Bladder Cancer

一个小鼠膀胱癌原位模型

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and ...

科研

医学

Orthotopic Small Bowel Transplantation in Rats

大鼠原位小肠移植

Small  bowel transplantation has become an accepted clinical option for patients with  short gut syndrome and failure of parenteral nutrition ...

Orthotopic Liver Transplantation in Rats

大鼠原位肝移植

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

Imaging Glioma Initiation In Vivo Through a Polished and Reinforced Thin-skull Cranial Window

Imaging Glioma Initiation In Vivo Through a Polished and Reinforced Thin-skull Cranial Window

成像神经胶质瘤启动<em在体内</em&gt;通过抛光和增强的薄膜头骨的颅窗

Glioma is the one of the most lethal forms of human cancer. The most effective glioma therapy to date-surgery followed by radiation treatment-offers ...

Fetal Echocardiography and Pulsed-wave Doppler Ultrasound in a Rabbit Model of Intrauterine Growth Restriction

在家兔模型宫内生长受限胎儿超声心动图和多普勒超声脉冲波

Fetal intrauterine growth restriction (IUGR) results in abnormal cardiac function that is apparent antenatally due to advances in fetoplacental Doppler ...

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

生物荧光原位模型胰腺癌进展

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%.  New therapeutic options are critically needed and depend on improved ...

Generation of Subcutaneous and Intrahepatic Human Hepatocellular Carcinoma Xenografts in Immunodeficient Mice

皮下及肝内人肝癌裸鼠移植瘤的免疫缺陷小鼠的产生

In  vivo experimental models of hepatocellular carcinoma (HCC) that recapitulate  the human disease provide a valuable platform for research into disease  ...

Minimally Invasive Establishment of Murine Orthotopic Bladder Xenografts

微创建立小鼠原位膀胱移植瘤的

Orthotopic bladder cancer xenografts are the gold standard to study molecular cellular manipulations and new therapeutic agents&#160;in vivo. Suitable ...

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 ...

Creating Anatomically Accurate and Reproducible Intracranial Xenografts of Human Brain Tumors

创建精确的解剖学和人脑肿瘤的重现性颅内移植瘤

Orthotopic tumor models are currently the best way to study the characteristics of a tumor type, with and without intervention, in the context of a live ...