Name: the in ovo cam-assay as a xenograft model for sarcoma
Uploaded: 2013-07-17T21:00:00
Description: Watch this Scientific Journal Video about The In ovo CAM-assay as a Xenograft Model for Sarcoma at JoVE.com

从SW1353软骨肉瘤细胞系细胞恳请教授的PCW Hogendoorn博士，荷兰莱顿大学教授J.博维。我们感谢的J. Mestach和G. Wagemans优秀的技术援助，G.德Bruyne专业绘图概述我们的协议。

参见图1的概述肿瘤材料 1。肿瘤样本的获取和准备的伦理委员会的批准，如果在病人的材料的使用是必要的，从病人取得的知情同意书。 <ol><li>嘉实代表材料（最小为1厘米3）干预的时间，无论是活检或切除的肉瘤。活检定义适当的地点使用动态对比度MRI。 </li><li>立即将材料在无菌小瓶含1000 U青霉素和1毫克/毫升?...

<ol>
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(33), e1620 (2009).</li><li>Balke, M., Neumann, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphologic+characterization+of+osteosarcoma+growth+on+the+chick+chorioallantoic+membrane.">Morphologic characterization of osteosarcoma growth on the chick chorioallantoic membrane.</a> BMC Research Notes. 3, 58 (2010).</li><li>Hendrix, A., Maynard, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+the+secretory+small+GTPase+Rab27B+on+breast+cancer+growth,+invasion,+and+metastasis.">Effect of the secretory small GTPase Rab27B on breast cancer growth, invasion, and metastasis.</a> Journal of the National Cancer Institute. 102, 866-880 (2010).</li><li>Ausprunk, D., Knighton, D., Folkman, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascularization+of+normal+and+neoplastic+tissues+grafted+to+the+chick+chorioallantois:+role+of+host+and+preexisting+graft+vessels.">Vascularization of normal and neoplastic tissues grafted to the chick chorioallantois: role of host and preexisting graft vessels.</a> American Journal of Pathology. 79, 597-618 (1975).</li><li>Lokman, N. 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接种和收获的时间时序进行SAOS2天接种ECM凝胶（36的CAM）和胚胎发育天5和10之间变化。 孵化前9天，CAM是不能持续大到足以支持我们采用ECM凝胶。收获时，肿瘤细胞有时不得不从更深的CAM检索，并在蛋白或CAM一些ECM凝胶样品躺在松动。在第5和第6天，这是很难避免把小鸡畸形或死亡，导致肿瘤细胞鸡胚。从第9天，在CAM被完全覆盖的大部分鸡蛋的窗口下面的表?...

肉瘤是一种罕见的肿瘤治疗抵抗1,2由于死亡率高的结缔组织。患者的生存率进展受阻低的年发病，其广泛的多样性，以及肉瘤细胞的事实，据报道， 在体外难以培养3,4。 使用培养细胞的临床前治疗评价已经表明，新的，显然是在体外的活性分子并不总是反映在临床上的结果。此外，揭示了基因表达阵列的基因畸变并不总是与个别病人5-7的肿瘤行为特征。为了试图解决这些问题，已经获得了个性化医疗的重要性，这也反映在增加搜索异种移植模型8-12。 在体内试验中的一种反映复杂的相互作用的优点是与c的相关关系在固体肿瘤细胞与宿主组织环境，必要的癌细胞增殖和侵袭13。目前，我们研究使用绒毛膜尿囊膜法（CAM法）作为一个可重复的异种移植物模型肉瘤14,15。本试剂盒被广泛用于肿瘤血管生成的研究16,17。然而，在文献中，我们已经发现这个实验的不同的协议，而其他的研究观察到的生长或血管生成的显着性差异，根据不同的协议18,19。 在这篇文章中，我们探讨不同的CAM检测条件对细胞行为的肿瘤移植，肿瘤来源的单细胞悬液和既定的肉瘤细胞培养的效果。

<table border="1">
	<tbody>
		<tr>
			<td>Name of Reagent</td>
			<td>Company</td>
			<td>Catalog Number</td>
		</tr>
		<tr>
			<td>Cell Line Nucleofector Kit V</td>
			<td>Amaxa</td>
			<td>VCA-1003</td>
		</tr>
		<tr>
			<td>collagenase 2 solution (500 U/ml RPMI 1640)</td>
			<td>Sigma Aldrich</td>
			<td>C6885</td>
		</tr>
		<tr>
			<td>DMEM</td>
			<td>Invitrogen</td>
			<td>41965-039</td>
		</tr>
		<tr>
			<td>DMSO</td>
			<td>Sigma</td>
			<td>D8418</td>
		</tr>
		<tr>
			<td>Dnase solution</td>
			<td>Sigma Aldrich</td>
			<td>DN25</td>
		</tr>
		<tr>
			<td>G418</td>
			<td>Invitrogen</td>
			<td>11811031</td>
		</tr>
		<tr>
			<td>Matrigel</td>
			<td>Sigma-Aldrich</td>
			<td>E1270</td>
		</tr>
		<tr>
			<td>mouse primary monoclonal antibody Ki67</td>
			<td>Dako Denmark</td>
			<td>MIB-1</td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>Fluka</td>
			<td>D76240</td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Invitrogen</td>
			<td>20012019</td>
		</tr>
		<tr>
			<td>PBSD</td>
			<td>Invitrogen</td>
			<td>14040083</td>
		</tr>
		<tr>
			<td>peGFP-C1 vector</td>
			<td>Clontech</td>
			<td>632470</td>
		</tr>
		<tr>
			<td>Penicillin/streptomycin</td>
			<td>Invitrogen</td>
			<td>15140163</td>
		</tr>
		<tr>
			<td>RPMI</td>
			<td>Invitrogen</td>
			<td>22409-015</td>
		</tr>
		<tr>
			<td>Trypsin-EDTA solution</td>
			<td>Invitrogen</td>
			<td>25300054</td>
		</tr>
		<tr>
			<td>Vybrant cell-labeling DiI</td>
			<td>Lifetechnologies</td>
			<td>22885</td>
		</tr>
		<tr>
			<td>Name of Equipment</td>
			<td>Company</td>
			<td>Catalog Number</td>
		</tr>
		<tr>
			<td>Countess Automated Cell Counter</td>
			<td>Invitrogen</td>
			<td>C10227</td>
		</tr>
		<tr>
			<td>digital color camera</td>
			<td>Leica</td>
			<td>DFC 340 FX</td>
		</tr>
		<tr>
			<td>Digital Egg Incubator</td>
			<td>Auto Elex Co</td>
			<td>R-COM 50</td>
		</tr>
		<tr>
			<td>FACS</td>
			<td>BD Biosciences</td>
			<td>FACSAriaIII</td>
		</tr>
		<tr>
			<td>Gentlemacs C-Tube</td>
			<td>Miltenyi Biotech</td>
			<td>130-093-237</td>
		</tr>
		<tr>
			<td>Gentlemacs Dissociator</td>
			<td>Miltenyi Biotech</td>
			<td>130-093-235</td>
		</tr>
		<tr>
			<td>Gentlemacs Dissociator User Manual containing h_tumor protocol</td>
			<td>Miltenyi Biotech</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>semipermeable adhesive film (Suprasorb F)</td>
			<td>Lohmann&amp;Rauscher</td>
			<td>20468</td>
		</tr>
		<tr>
			<td>stereo fluorescence microscope</td>
			<td>Leica</td>
			<td>M205 FA</td>
		</tr>
		<tr>
			<td>Tissue-Tek Film automated Coverslipper</td>
			<td>Sakura</td>
			<td>6400</td>
		</tr>
		<tr>
			<td>ultraView Universal DAB Detection Kit</td>
			<td>Ventana Medical Systems Inc</td>
			<td>760-500</td>
		</tr>
		<tr>
			<td>Ventana Automated Slide Stainer</td>
			<td>Ventana Medical Systems</td>
			<td>Benchmark XT</td>
		</tr>
	</tbody>
</table>

the in ovo cam-assay as a xenograft model for sarcoma

肉瘤是一种非常罕见的疾病，在本质上是异质的，都阻碍了发展的新疗法。肉瘤患者的理想人选后分层的个性化医学，解释目前在开发这种疾病的一个重现性好，成本低的异种移植模型的兴趣。鸡胚绒毛尿囊膜，是一种天然的免疫缺陷宿主能够维持嫁接的组织和细胞，没有种属特异性的限制。此外，它很容易访问，操纵和使用光学和荧光立体显微镜成像。组织学进一步允许异型细胞相互作用的详细分析。 该协议详细描述了OVO绒毛尿囊膜移植肉瘤派生的新鲜肿瘤组织，其单细胞悬液，和永久和瞬态荧光标记的建立肉瘤细胞株（系Saos-2和SW1353）。小鸡存活大鼠上课是高达75％。该模型用于研究移植（活力，Ki67的增殖指数，坏死，浸润）和主机（成纤维细胞浸润，血管长入）的行为。对于单细胞悬液的本地化嫁接，ECM凝胶提供了显着的优势，在惰性容器材料。 Ki67增殖指数的距离的凸轮表面的细胞和CAM上，后者确定的治疗产品的另外的时间框架中的应用的持续时间有关。

在CAM的评价肿瘤移植变得附着到CAM（ 图2A）。单细胞悬液经常从病人的材料显示干燥，微微凸起的牌匾（ 图2D）。切除后的CAM，标志着起皱发生膜（ 图2E和2F）。 对于商业细胞株的牌匾变得更加不透明，表明细胞的增殖。 ECM凝胶在不同的细胞系有明显的增长模式上的CAM。 SAOS2形成一个蔓延斑块，表面从接?...

Watch this Scientific Journal Video about The In ovo CAM-assay as a Xenograft Model for Sarcoma at JoVE.com

The In ovo CAM-assay as a Xenograft Model for Sarcoma

该<em&gt;在OVO</em&gt;绒毛尿囊膜（CAM）接枝源性肉瘤的新鲜肿瘤组织中，其单细胞悬液，并永久和瞬态荧光标记的建立肉瘤细胞株。该模型用于研究移植（活力，Ki67的增殖指数，坏死，浸润）和主机（成纤维细胞浸润，血管长入）的行为。

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医学

A Simple Guide Screw Method for Intracranial Xenograft Studies in Mice

一个简单的指南螺杆颅内移植瘤小鼠的研究方法

The grafting of human tumor cells into the brain of immunosuppressed mice is an established method for the study of brain cancers including glioblastoma ...

Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy

人类神经内分泌肿瘤细胞株为研究人类神经内分泌肿瘤治疗的三维模型

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human ...

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

脑Microslices的兴奋性刺激作为<em&gt;在体外</em&gt;中风模型

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As ...

Glutamate and Hypoxia as a Stress Model for the Isolated Perfused Vertebrate Retina

谷氨酸和缺氧作为离体脊椎动物视网膜应力模型

Neuroprotection has been a strong field of investigation in ophthalmological research in the past decades and affects diseases such as glaucoma, retinal ...

The In Ovo Chick Chorioallantoic Membrane (CAM) Assay as an Efficient Xenograft Model of Hepatocellular Carcinoma

The In Ovo Chick Chorioallantoic Membrane CAM Assay as an Efficient Xenograft Model of Hepatocellular Carcinoma

该<em&gt;卵内</em&gt;鸡胚尿囊膜（CAM）含量为肝癌的有效异种移植模型

The chick chorioallantoic membrane (CAM) begins to develop by day 7 after fertilization and matures by day 12. The CAM is naturally immunodeficient and ...

Establishment of a Human Multiple Myeloma Xenograft Model in the Chicken to Study Tumor Growth, Invasion and Angiogenesis

建立一个人类多发性骨髓瘤异种移植模型中的鸡，研究肿瘤的生长，侵袭和血管生成

Multiple myeloma (MM), a malignant plasma cell disease, remains incurable and novel drugs are required to improve the prognosis of patients. Due to the ...

Oleic Acid-Injection in Pigs As a Model for Acute Respiratory Distress Syndrome

油酸注射液在猪急性呼吸窘迫综合征模型中的研究

The acute respiratory distress syndrome is a relevant intensive care disease with an incidence ranging between 2.2% and 19% of intensive care unit ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing

眼毛刺角膜上皮磨损与角膜创面愈合的模型研究

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to ...