Name: intracranial orthotopic allografting of medulloblastoma cells in immunocompromised mice
Uploaded: 2010-10-03T17:00:00
Description: Watch this Scientific Journal Video about Intracranial Orthotopic Allografting of Medulloblastoma Cells in Immunocompromised Mice at JoVE.com

这项研究是由Vanderbilt - Ingram癌症中心支援津贴（P30 CA068485），儿童脑瘤基金会和美国国立卫生研究院（NS042205）的赠款支持。

1。荷瘤小脑和肿瘤组织中的分离的显微解剖<ol><li> 检索肿瘤组织 <ol><li>轴承髓母细胞瘤的患病老鼠往往runted，并显示脑积水和典型的神经症状，包括瘫痪后未能收复当朝天的姿势。要检索肿瘤组织，安乐死小鼠吸入二氧化碳。这是项重要的，不执行颈椎脱位，程序产生的压力后的头骨，并能破坏肿瘤组织的完整性。 </li><li>斩首是死亡后，立即用一把剪刀，头发和肌肉组织，尽可能...

<ol>
	<li>Schuller, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acquisition+of+granule+neuron+precursor+identity+is+a+critical+determinant+of+progenitor+cell+competence+to+form+Shh-induced+medulloblastoma.">Acquisition of granule neuron precursor identity is a critical determinant of progenitor cell competence to form Shh-induced medulloblastoma.</a> Cancer Cell. 14, 123-134 (2008).</li><li>Yang, Z. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Medulloblastoma+can+be+initiated+by+deletion+of+Patched+in+lineage-restricted+progenitors+or+stem+cells.">Medulloblastoma can be initiated by deletion of Patched in lineage-restricted progenitors or stem cells.</a> Cancer Cell. 14, 135-145 (2008).</li><li>Kessler, 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=N-myc+alters+the+fate+of+preneoplastic+cells+in+a+mouse+model+of+medulloblastoma.">N-myc alters the fate of preneoplastic cells in a mouse model of medulloblastoma.</a> Genes Dev. 23, 157-170 (2009).</li><li>Flora, A., Klisch, T. J., Schuster, G., Zoghbi, H. Y., Y, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deletion+of+Atoh1+disrupts+Sonic+Hedgehog+signaling+in+the+developing+cerebellum+and+prevents+medulloblastoma.">Deletion of Atoh1 disrupts Sonic Hedgehog signaling in the developing cerebellum and prevents medulloblastoma.</a> Science. 326, 1424-1427 (2009).</li><li>Salsano, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+the+neurogenic+basic+helix-loop-helix+transcription+factor+NEUROG1+identifies+a+subgroup+of+medulloblastomas+not+expressing+ATOH1.">Expression of the neurogenic basic helix-loop-helix transcription factor NEUROG1 identifies a subgroup of medulloblastomas not expressing ATOH1.</a> Neuro Oncol. 9, 298-307 (2007).</li></ol>

几个关键因素，以确保成功的髓母细胞瘤细胞allografting，产生继发性肿瘤的形成如下：首先，用于组织解离，只有50％Accutase和不过消化的组织，作为长期的酶处理细胞活力显着降低。另外，只使用1毫升的大小Pipetman较小的提示，还可以生成游离细胞的物理损坏。它是好的，有单个细胞和小allografting聚集的混合物。二，混合和平衡的肿瘤细胞的解决方案，每次装货前汉密尔顿注射器。不要使用超过...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>Neurobasal medium</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>21103049</td>
<td>&nbsp;</td>
<tr>
<td>hEGF</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>PHG0311</td>
<td>25ng/ml</td>
</tr>
<tr>
<td>bFGF</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>PHG0023</td>
<td>25ng/ml</td>
</tr>
<tr>
<td>N2</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>17502048</td>
<td>1X</td>
</tr>
<tr>
<td>B27(-RA)</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>12587010</td>
<td>1X</td>
</tr>
<tr>
<td>Accutase</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>A1110501</td>
<td>50% </td>
</tr>
<tr>
<td>Glutamine</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>25030081</td>
<td>2mM</td>
</tr>
<tr>
<td>Stereotaxic instrument</td>
<td>&nbsp;</td>
<td>Stoelting </td>
<td>51730</td>
<td>&nbsp;</td>
<tr>
<td>Foredom flexible shaft drill, hang-up style</td>
<td>&nbsp;</td>
<td>Stoelting </td>
<td>58650</td>
<td>&nbsp;</td>
<tr>
<td>Large probe holder</td>
<td>&nbsp;</td>
<td>Stoelting </td>
<td>51633</td>
<td>&nbsp;</td>
<tr>
<td>10 &mu;L syringe, 26 ga., bevel tip</td>
<td>&nbsp;</td>
<td>Stoelting </td>
<td>51105</td>
<td>&nbsp;</td>
<tr>
<td>Drill bit .75mm</td>
<td>&nbsp;</td>
<td>Stoelting</td>
<td>51455-3</td>
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intracranial orthotopic allografting of medulloblastoma cells in immunocompromised mice

髓母细胞瘤是最常见的小儿神经系统的肿瘤。小脑颗粒神经元前体（CGNPs）1-4为髓母细胞瘤细 ​​胞的原产地集中于动物研究的一个庞大的身躯。然而，在人类患者（结节，促纤维增生，古典和大细胞/间变性），而事实上，在没有CGNP标记5的异位表达的人类患者的一个子集发现髓母细胞瘤是，髓母细胞瘤亚型的不同临床表现表明，髓母细胞瘤细胞和分子的起源更复杂，远未完全破译。因此，它是必不可少的，以确定是否有替代髓母细胞瘤肿瘤细胞的起源细胞类型，具体的治疗方法可以开发基于。为此，颅内原位基因标记的肿瘤细胞类型肿瘤的发展中，收件人的后续分析allografting将使测定肿瘤细胞的细胞来源。在这里，我们描述了颅内原位allografting髓母细胞瘤细胞从原发肿瘤组织的实验协议，这个过程也可以用于移植建立细胞株的细胞。

Watch this Scientific Journal Video about Intracranial Orthotopic Allografting of Medulloblastoma Cells in Immunocompromised Mice at JoVE.com

Intracranial Orthotopic Allografting of Medulloblastoma Cells in Immunocompromised Mice

这个协议描述鼠标髓母细胞瘤组织的隔离和分裂，并随后allografting到免疫功能低下的受体小鼠的肿瘤细胞，以开展二次髓母细胞瘤。

在免疫功能低下小鼠颅内髓母细胞瘤细胞原位Allografting

Medulloblastoma is the most common pediatric tumor of the nervous system. A large body of animal studies has focused on cerebellar granule neuron ...

Research

JoVE Journal

Neuroscience

Isolation, Enrichment, and Maintenance of Medulloblastoma Stem Cells

髓母细胞瘤干细胞的分离，浓缩，和维修

Brain tumors have been suggested to possess a small population of stem cells that are the root cause of tumorigenesis. Neurosphere assays have been ...

科研

神经科学

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在完整的感觉器官前体细胞的活细胞成像果蝇蛹

Since the discovery of Green Fluorescent Protein (GFP), there has been a revolutionary change in the use of live-cell imaging as a tool for understanding ...

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

测定冷灵敏度和适应小鼠一种简单廉价的方法

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

Intracranial Pharmacotherapy and Pain Assays in Rodents

颅内药物治疗和啮齿动物疼痛检测

Pain is a salient sensory experience with affective and cognitive dimensions. However, central mechanisms for pain remain poorly understood, hindering the ...

The Detection of 5-Hydroxymethylcytosine in Neural Stem Cells and Brains of Mice

小鼠神经干细胞和脑中5-羟基甲基细胞氨酸的检测

Multiple&#160;DNA modifications have been identified in the mammalian genome. Of that, 5-methylcytosine and 5-hydroxymethylcytosine-mediated epigenetic ...

Quantitative Measurement of Intrathecally Synthesized Proteins in Mice

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Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the ...

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使用腺相关病毒在小鼠中瞄准科菲林操纵的大脑皮层区域的电皮质记录

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