Name: experimental metastasis assay
Uploaded: 2010-08-24T17:10:00
Description: Watch this Scientific Journal Video about Experimental Metastasis Assay at JoVE.com

このプロトコルは、検査の結果、博士リチャードハインズ（MIT）の研究室で最初に最適化されていました。資金調達はNYSTEMアイデア賞（LXへ）とルースL.キルシュシュタイン国立リサーチサービス賞（LXへの）によって提供されます。

1。サンプル準備<ol><li>成長因子やFBS（例えば、MC - 1細胞​​を10％FBS含有DMEM）で、その特定のメディアで〜70％コンフルエントに細胞を成長させる。プレートから培地を吸引除去し、穏やかに1 × PBS（塩化ナトリウム8gの/ L、塩化カリウム0.2 g / Lの、1.15のNa 2 HPOのG / L 4 · 7H 2 O、KH 2 POを0.2g / Lで数回洗浄4、pHは7.3）。 </li><li>吸引PBSとヴェルセンの0.05?...

<ol>
	<li>Xu, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+expression+changes+in+an+animal+melanoma+model+correlate+with+aggressiveness+of+human+melanoma+metastases.">Gene expression changes in an animal melanoma model correlate with aggressiveness of human melanoma metastases.</a> Mol Cancer Res. 6 (5), 760-760 (2008).</li><li>Clark, E. A., Golub, T. R., Lander, E. S., Hynes, R. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genomic+analysis+of+metastasis+reveals+an+essential+role+for+RhoC.">Genomic analysis of metastasis reveals an essential role for RhoC.</a> Nature. 406 (6795), 532-532 (2000).</li><li>Xu, L., Begum, S., Hearn, J. D., Hynes, R. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=GPR56,+an+atypical+G+protein-coupled+receptor,+binds+tissue+transglutaminase,+TG2,+and+inhibits+melanoma+tumor+growth+and+metastasis.">GPR56, an atypical G protein-coupled receptor, binds tissue transglutaminase, TG2, and inhibits melanoma tumor growth and metastasis.</a> Proc Natl Acad Sci U S A. 103 (24), 9023-9023 (2006).</li><li>Kozlowski, J. M., Hart, I. R., Fidler, I. J., Hanna, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+human+melanoma+line+heterogeneous+with+respect+to+metastatic+capacity+in+athymic+nude+mice.">A human melanoma line heterogeneous with respect to metastatic capacity in athymic nude mice.</a> J Natl Cancer Inst. 72 (4), 913-913 (1984).</li><li>Kang, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+multigenic+program+mediating+breast+cancer+metastasis+to+bone.">A multigenic program mediating breast cancer metastasis to bone.</a> Cancer Cell. 3 (6), 537-537 (2003).</li></ol>

転移は、がん患者における死亡の主要な原因である。彼らの主要な遺伝子座、循環への参入（管内）、循環（血管外漏出）から、これらの終了、および遠隔臓器の生存と成長から癌細胞の剥離：これは、4つの主要な手順が含まれます。人間の転移はゆっくりとしたプロセスとみなされ、多くの場合、待ち時間の年後に明示​​される。タイムリーにその進行を勉強するために、上記の比較的...

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		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>DMEM</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>10313-021</td>
<td>&nbsp;</td>
<tr>
<td>FBS</td>
<td>&nbsp;</td>
<td>Hyclone</td>
<td>SH30088.03</td>
<td>&nbsp;</td>
<tr>
<td>HBSS</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>14175-095</td>
<td>&nbsp;</td>
<tr>
<td>NaCl</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>S9888</td>
<td>&nbsp;</td>
<tr>
<td>KCl</td>
<td>&nbsp;</td>
<td>Mallinckrodt Chemicals</td>
<td>6838-04</td>
<td>&nbsp;</td>
<tr>
<td>Na2 HPO4.7H2O</td>
<td>&nbsp;</td>
<td>Mallinckrodt Chemicals</td>
<td>7896-04</td>
<td>&nbsp;</td>
<tr>
<td>KH2PO4</td>
<td>&nbsp;</td>
<td>Mallinckrodt Chemicals</td>
<td>7100-12</td>
<td>&nbsp;</td>
<tr>
<td>Phenol red</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>P3532</td>
<td>&nbsp;</td>
<tr>
<td>EDTA-Na</td>
<td>&nbsp;</td>
<td>EMD Biosciences</td>
<td>4010</td>
<td>&nbsp;</td>
<tr>
<td>Trypan Blue</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>15250-061</td>
<td>&nbsp;</td>
<tr>
<td>Trypsin 2.5%</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>15090-046</td>
<td>&nbsp;</td>
<tr>
<td>Ethanol</td>
<td>&nbsp;</td>
<td>Ultrapure</td>
<td>200-CSGP</td>
<td>&nbsp;</td>
<tr>
<td>50 ml Falcon Tube</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>89039-656</td>
<td>&nbsp;</td>
<tr>
<td>Microcentrifuge Tubes</td>
<td>&nbsp;</td>
<td>Axygen Scientific</td>
<td>MCT-175-C</td>
<td>&nbsp;</td>
<tr>
<td>Falcon 70&#181;m Cell Strainer</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>352350</td>
<td>&nbsp;</td>
<tr>
<td>5ml Falcon Culture Tube</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>60818-576</td>
<td>&nbsp;</td>
<tr>
<td>Hemacytometer</td>
<td>&nbsp;</td>
<td>Hausser Scientific</td>
<td>15170-208</td>
<td>&nbsp;</td>
<tr>
<td>1 ml syringe</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>329650</td>
<td>&nbsp;</td>
<tr>
<td>30 &frac12; gauge needle</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>305106</td>
<td>&nbsp;</td>
<tr>
<td>Mouse restrainer</td>
<td>&nbsp;</td>
<td>Plas Labs Inc.</td>
<td>551-BSRR</td>
<td>&nbsp;</td>
<tr>
<td>Surgical scissors</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>08-940</td>
<td>&nbsp;</td>
<tr>
<td>Forceps</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>08-902</td>
<td>&nbsp;</td>
<tr>
<td>Dissecting Microscope</td>
<td>&nbsp;</td>
<td>Nikon</td>
<td>SMZ1500</td>
<td>&nbsp;</td>
<tr>
<td>Benchtop Centrifuge</td>
<td>&nbsp;</td>
<td>Thermoscientific</td>
<td>75003491</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

experimental metastasis assay

転移は、がん患者における死亡の主要な原因である。転移のメカニズムを理解するために、実験的転移アッセイは、免疫​​不全マウスを用いて設立されました。この記事では、サンプル調製、静脈内注射、および肺転移から培養細胞を含むこの検定に関わる手順を、群を区別。簡単に言えば、ヒト癌細胞の予め決められた数を調製した<em&gt; in vitroで</em&gt;とは直接彼らの尾静脈を介して免疫不全マウスの循環に注入。細胞の数が少ない循環の乱後も存続し、肺などの臓器、の転移として成長する。注入されたマウスは、一定期間後に解剖されています。転移の組織分布は、解剖顕微鏡下で決定されます。特定の組織への転移の数がカウントされ、それが直接注入された癌細胞の転移能と相関する。生じた転移を単離し、培養する<em&gt; in vitroで</em&gt;のような免疫不全マウスに再び注入すると、しばしば親系統よりも強化された転移性の能力を示す細胞株、。これらの転移性の高いデリバティブ取引は、転移性の進行を調節する遺伝子や分子経路を同定するために有用なツールとなる。

Watch this Scientific Journal Video about Experimental Metastasis Assay at JoVE.com

Experimental Metastasis Assay

この記事では、ヒト癌細胞株の転移能を決定するために使用される実験的転移アッセイの手順を説明します。

実験的転移アッセイ

Metastasis is the leading cause of death in cancer patients.  To understand the mechanism of metastasis, an experimental metastasis assay was established ...

Research

JoVE Journal

Medicine

Quantitative Analysis of Cancer Metastasis using an Avian Embryo Model

鳥類胚のモデルを使用して、がんの転移の定量的解析

During metastasis cancer cells disseminate from the primary tumor, invade into surrounding tissues, and spread to distant organs. Metastasis is a complex ...

In vitro Mesothelial Clearance Assay that Models the Early Steps of Ovarian Cancer Metastasis

In vitro Mesothelial Clearance Assay that Models the Early Steps of Ovarian Cancer Metastasis

in vitroで中皮クリアランスアッセイでそのモデルの卵巣がんの転移の初期段階を

Ovarian cancer is the fifth leading cause of cancer related deaths in the United States1. Despite a positive initial response to therapies, 70 to 90 ...

Models of Bone Metastasis

骨転移モデル

Bone metastases are a common occurrence in several malignancies, including breast, prostate, and lung. Once established in bone, tumors are responsible ...

Experimental Human Pneumococcal Carriage

実験的な人間の肺炎球菌キャリッジ

Experimental human pneumococcal carriage (EHPC) is scientifically important because nasopharyngeal carriage of Streptococcus pneumoniae is both the major ...

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

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

ザ<em&gt;卵で</em肉腫のためXenograftmodelとして&gt; CAMアッセイ

Sarcoma is a very rare disease that is heterogeneous in nature, all hampering the development of new therapies. Sarcoma patients are ideal candidates for ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

ヒト前立腺癌転移の同所マウスモデル

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

Primary Tumor and MEF Cell Isolation to Study Lung Metastasis

原発腫瘍と肺転移を研究するためのMEF細胞の単離

In breast tumorigenesis, the metastatic stage of the disease poses the greatest threat to the affected individual. Normal breast cells with altered ...

Chick Heart Invasion Assay for Testing the Invasiveness of Cancer Cells and the Activity of Potentially Anti-invasive Compounds

テストのためのひよこ心臓浸潤アッセイ癌細胞の浸潤性および潜在的に抗浸潤化合物の活性

The goal of the chick heart assay is to offer a relevant organ culture method to study tumor invasion in three dimensions. The assay can distinguish ...

A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

肝細胞癌転移の「患者様」同所同系マウスモデル

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of ...

Quantitation of Intra-peritoneal Ovarian Cancer Metastasis

腹腔内卵巣癌転移の定量

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States.  Mortality is due to diagnosis of 75% of ...