Name: modeling and imaging 3-dimensional collective cell invasion
Uploaded: 2011-12-07T13:00:00
Description: Watch this Scientific Journal Video about Modeling and Imaging 3-Dimensional Collective Cell Invasion at JoVE.com

资助这项研究是由英国癌症研究中心。

1。逆转录病毒荧光蛋白标记的细胞<ol><li>板逆转录病毒包装细胞（ 如凤凰卫视）出0.25 × 10 6细胞，一个6孔，每孔 菜在10％胎牛血清（FBS）/ DMEM培养液。 </li><li>使用逆转录病毒DNA转染细胞48小时后细胞Effectene根据制造商的指示。 </li><li>培养24小时冲洗井两次后，再加入1.5毫升10％FBS /每口井的DMEM。 </li><li>收集包装病毒在组织培养中等48小时后吹打，并转移至2 mL离心管?...

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	<li>Olson, M. F., Sahai, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+actin+cytoskeleton+in+cancer+cell+motility.">The actin cytoskeleton in cancer cell motility.</a> Clin. Exp. Metastasis. 26, 273-287 (2008).</li><li>Hoelzinger, D. B., Demuth, T., Berens, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autocrine+factors+that+sustain+glioma+invasion+and+paracrine+biology+in+the+brain+microenvironment.">Autocrine factors that sustain glioma invasion and paracrine biology in the brain microenvironment.</a> J. Natl. Cancer. Inst. 99, 1583-1593 (2007).</li><li>Chaffer, C. L., Weinberg, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+perspective+on+cancer+cell+metastasis.">A perspective on cancer cell metastasis.</a> Science. 331, 1559-1564 (2011).</li><li>Hanahan, D., Weinberg, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Hallmarks+of+Cancer.">The Hallmarks of Cancer.</a> Cell. 100, 57-70 (2000).</li><li>Francia, G., Cruz-Munoz, W., Man, S., Xu, P., Kerbel, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+advanced+spontaneous+metastasis+for+experimental+therapeutics.">Mouse models of advanced spontaneous metastasis for experimental therapeutics.</a> Nat. Rev. Cancer. 11, 135-141 (2011).</li><li>Hooper, S., Marshall, J. F., Sahai, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+cell+migration+in+three+dimensions.">Tumor cell migration in three dimensions.</a> Methods. Enzymol. 406, 625-643 (2006).</li><li>Croft, D. R., Olson, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulating+the+conversion+between+rounded+and+elongated+modes+of+cancer+cell+movement.">Regulating the conversion between rounded and elongated modes of cancer cell movement.</a> Cancer Cell. 14, 349-351 (2008).</li><li>Wolf, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multi-step+pericellular+proteolysis+controls+the+transition+from+individual+to+collective+cancer+cell+invasion.">Multi-step pericellular proteolysis controls the transition from individual to collective cancer cell invasion.</a> Nat. Cell. Biol. 9, 893-904 (2007).</li><li>Hennigan, R. F., Hawker, K. L., Ozanne, B. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fos-transformation+activates+genes+associated+with+invasion.">Fos-transformation activates genes associated with invasion.</a> Oncogene. 9, 3591-3600 (1994).</li><li>Scott, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=LIM+kinases+are+required+for+invasive+path+generation+by+tumor+and+tumor-associated+stromal+cells.">LIM kinases are required for invasive path generation by tumor and tumor-associated stromal cells.</a> J. Cell. Biol. 191, 169-185 (2010).</li></ol>

基质胶侵袭实验历来设立细胞到细胞外基质蛋白与趋化因子诱导的蠕动层对通过底部的过滤器放在。侵袭得分多少细胞可以在过滤器的底面计算的函数。虽然实际上有以上所述的“逆”入侵检测的差别不大，有相当多的入侵过程中，可以通过可视化入侵的细胞，因为他们通过基质胶和确定的信息。例如，除了能够看到不同的标记细胞研究领导与集体入侵细胞（图3C和3D），使用这种方法使细胞固定...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th></tr><tr><td> DMEM（贝科改良Eagle培养基） </td><td> GIBCO公司 </td><td> 21969 </td></tr><tr><td>胎牛血清</td><td> 临时机场管理局 </td><td> A15 - 101 </td></tr><tr><td>青霉素链霉素</td><td> GIBCO公司 </td><td> 15140 </td></tr><tr><td> 200毫米L -谷氨酰胺（100X） </td><td> GIBCO公司 </td><td> 25050-032 </td></tr><tr><td>嘌呤霉素</td><td> Sigma - Aldrich公司 </td><td> P8833 </td></tr><tr><td> 0.05％胰蛋白酶EDTA </td><td> GIBCO公司 </td><td> 25300 </td></tr><tr><td>凝聚胺</td><td> Sigma - Aldrich公司 </td><td> AL - 118 </td></tr><tr><td>脂质体2000试剂</td><td> Invitrogen公司 </td><td> 11668019 </td></tr><tr><td> 6.5毫米Transwells，8.0微米孔径</td><td> 康宁 </td><td> 3422 </td></tr><tr><td>完整的基质胶</td><td> BD公司 </td><td> 354234 </td></tr> &lt;TR&gt; <td>钙黄绿素AM </td><td> Invitrogen公司 </td><td> C1430 </td></tr><tr><td>核糖核酸酶</td><td> Qiagen公司 </td><td> 19101 </td></tr><tr><td>碘化丙啶</td><td> Sigma - Aldrich公司 </td><td> P4864 </td></tr><tr><td>共焦microcope </td><td> 徕卡 </td><td> SP2MP </td></tr></tbody></table>

modeling and imaging 3-dimensional collective cell invasion

一个定义特征是癌症的恶性肿瘤侵袭和转移1。在某些癌症（E. 克 。胶质瘤2），局部浸润到周围的健康组织的疾病和死亡的根源。其他癌症（E. 克 。乳腺癌，肺癌等 ），它是在转移过程中，肿瘤细胞从原发肿瘤的质量，殖民远端站点，并最终有助于器官功能衰竭，最终导致发病率和死亡率3。据估计，侵袭和转移是负责90％的癌症死亡 4 。因此，有浓厚的兴趣，提高诊断和治疗5的目的确定的分子过程，侵袭和转移的关键蛋白质调解员。 癌症科学家面临的一个挑战是要发展入侵检测， 充分类似于在体内的情况启用准确的疾病建模 6 。二维细胞活力检测信息约一个入侵方面，并没有考虑到帐户外基质（ECM）蛋白的重塑，这也是一个关键因素。最近，研究完善我们了解肿瘤细胞的侵袭，并透露，单个细胞的移动拉长或圆形模式 7 。此外，有更大的集体入侵的贡献表示赞赏，特别是在高度分化的肿瘤，维持上皮细胞的特点，使细胞股，床单和集群入侵 ，8癌症的蔓延。 我们目前的候选蛋白的研究贡献的集体入侵10改进后的方法9。特别是在单独池工程细胞表达不同的荧光蛋白，它是分子剖析的活动和proteiNS需要在领导与那些需要在以下细胞的细胞。使用的RNAi实验拆卸单个细胞的侵袭，以及在不同岗位的集体入侵所涉及的过程提供了分子工具。在此过程中，荧光标记的细胞的混合物镀以前与基质胶流脑蛋白填补了Transwell小插入底部，然后让其通过过滤器进入基质胶入侵的“向上”。 Z系列的形象栈，通过共聚焦成像到三维表示，重建允许集体入侵链分析和荧光标记的细胞，在领导与以下职位的任职可视化。

Watch this Scientific Journal Video about Modeling and Imaging 3-Dimensional Collective Cell Invasion at JoVE.com

Modeling and Imaging 3-Dimensional Collective Cell Invasion

成三维的细胞外基质肿瘤细胞的侵袭模型更好地反映<em&gt;在体内</em&gt;比二维的蠕动检测情况。可使用矩阵入侵检测结合共焦成像与荧光标记的细胞，对领导与下面的细胞入侵模式和独特的贡献的详细信息。

3维建模和成像集体细胞的侵袭

A defining characteristic of cancer malignancy is invasion and metastasis 1. In some cancers (e.g. glioma 2), local invasion into surrounding healthy ...

Research

JoVE Journal

Medicine

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

多光子成像在口腔鳞状细胞癌原位小鼠模型的肿瘤细胞的侵袭

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

科研

医学

Spheroid Assay to Measure TGF-&#946;-induced Invasion

Spheroid Assay to Measure TGF-ÃƒÅ½Ã‚Â²-induced Invasion

球体的检测，以测量TGF -β诱导的入侵

TGF-&beta; has opposing roles in breast cancer progression by acting as a tumor suppressor in the initial phase, but stimulating invasion and metastasis ...

3-Dimensional Resin Casting and Imaging of Mouse Portal Vein or Intrahepatic Bile Duct System

树脂浇铸和3维成像的小鼠门静脉或肝内胆管系统

In  organs, the correct architecture of vascular and ductal structures is  indispensable for proper physiological function, and the formation and  ...

Molecular Imaging to Target Transplanted Muscle Progenitor Cells

目标移植的肌肉祖细胞的分子成像

Duchenne muscular dystrophy (DMD) is a severe genetic neuromuscular disorder that affects 1 in 3,500 boys, and is characterized by progressive muscle ...

Generation and 3-Dimensional Quantitation of Arterial Lesions in Mice Using Optical Projection Tomography

生成和动脉病变的小鼠的三维定量使用光学投影层析成像

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating ...

Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and Ex vivo Fibroblasts

Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and Ex vivo Fibroblasts

在结直肠癌细胞的三维模型的入侵肿瘤微环境的分子剖析和<em&gt;体外</em&gt;成纤维细胞

Invading colorectal cancer (CRC) cells have acquired the capacity to break free from their sister cells, infiltrate the stroma, and remodel the ...

A Modified In vitro Invasion Assay to Determine the Potential Role of Hormones, Cytokines and/or Growth Factors in Mediating Cancer Cell Invasion

A Modified In vitro Invasion Assay to Determine the Potential Role of Hormones, Cytokines and/or Growth Factors in Mediating Cancer Cell Invasion

一种改进<em&gt;在体外</em&gt;入侵检测，以确定激素，在调节肿瘤细胞侵袭的细胞因子和/或生长因子的潜在作用

Blood serum serves as a chemoattractant towards which cancer cells migrate and invade, facilitating their intravasation into microvessels. However, the ...

Modeling Spontaneous Metastatic Renal Cell Carcinoma (mRCC) in Mice Following Nephrectomy

Modeling Spontaneous Metastatic Renal Cell Carcinoma mRCC in Mice Following Nephrectomy

在小鼠模型自发转移性肾细胞癌（MRCC）肾切除

One of the key challenges to improved testing of new experimental therapeutics in renal cell carcinoma (RCC) is the development of models that faithfully ...

Isolation and Immortalization of Patient-derived Cell Lines from Muscle Biopsy for Disease Modeling

The generation of patient-specific cell lines represents an invaluable tool for diagnostic or translational research, and these cells can be collected ...

A Cancer Cell Spheroid Assay to Assess Invasion in a 3D Setting

癌细胞球体检测，以评估入侵的3D设置

The invasive nature of cancer cell lines is thought to correlate with their metastatic potential.  Most traditional assays, however, do not examine these ...