Name: quantitative analysis of cancer metastasis using an avian embryo model
Uploaded: 2011-05-30T22:00:00
Description: Watch this Scientific Journal Video about Quantitative Analysis of Cancer Metastasis using an Avian Embryo Model at JoVE.com

हम उदारता से निषेचित अंडे प्रदान करने के लिए टायसन फूड्स इंक पहचान करना चाहते हैं. Shanna अर्नोल्ड इस प्रोटोकॉल के फिल्मांकन के दौरान महत्वपूर्ण भूमिका निभाई थी. Trenis पामर रुथ एल Kirschstein राष्ट्रीय अनुसंधान सेवा पुरस्कार (F31 राष्ट्रीय कैंसर संस्थान) के तहत स्वास्थ्य के राष्ट्रीय संस्थान द्वारा समर्थित किया गया. इस काम आंशिक रूप से CCSRI # 700537 JDL और NIH / NCI अनुदान # CA120711-01A1-Z CA120711-01A1 अनुदान द्वारा समर्थित किया गया था.

1. Xenografting के लिए अंडे (सहज मेटास्टेसिस) की तैयारी <ol><li> हौसले रखी निषेचित चिकन अंडे एक rotating इनक्यूबेटर में 100 ° एफ पर 10 दिन और 60% आर्द्रता के लिए incubated हैं. अंडे हर घंटे में चार बार घुमाया जाता है. </li><li> विकास के 10 दिन मे?...

<ol>
	<li>Ghiso, A. g. u. i. r. r. e., Kovalski, J. A., K, ., Ossowski, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+dormancy+induced+by+downregulation+of+urokinase+receptor+in+human+carcinoma+involves+integrin+and+MAPK+signaling.">Tumor dormancy induced by downregulation of urokinase receptor in human carcinoma involves integrin and MAPK signaling.</a> J Cell Biol. 147, 89-104 (1999).</li><li>Chambers, A. F., Shafir, R., Ling, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+model+system+for+studying+metastasis+using+the+embryonic+chick.">A model system for studying metastasis using the embryonic chick.</a> Cancer research. 42, 4018-4025 (1982).</li><li>Chambers, A. F., Wilson, S. M., Tuck, A. B., Denhardt, G. H., Cairncross, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+metastatic+properties+of+a+variety+of+mouse,+rat,+and+human+cells+in+assays+in+nude+mice+and+chick+embryos.">Comparison of metastatic properties of a variety of mouse, rat, and human cells in assays in nude mice and chick embryos.</a> In Vivo. 4, 215-219 (1990).</li><li>Deryugina, E. I., Zijlstra, A., Partridge, J. J., Kupriyanova, T. A., Madsen, M. A., Papagiannakopoulos, T., Quigley, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unexpected+effect+of+matrix+metalloproteinase+down-regulation+on+vascular+intravasation+and+metastasis+of+human+fibrosarcoma+cells+selected+in+vivo+for+high+rates+of+dissemination.">Unexpected effect of matrix metalloproteinase down-regulation on vascular intravasation and metastasis of human fibrosarcoma cells selected in vivo for high rates of dissemination.</a> Cancer Res. 65, 10959-10969 (2005).</li><li>Hahn-Dantona, E., Ramos-DeSimone, N., Sipley, J., Nagase, H., French, D. L., Quigley, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activation+of+proMMP-9+by+a+plasmin/MMP-3+cascade+in+a+tumor+cell+model.+Regulation+by+tissue+inhibitors+of+metalloproteinases.">Activation of proMMP-9 by a plasmin/MMP-3 cascade in a tumor cell model. Regulation by tissue inhibitors of metalloproteinases.</a> Ann N Y Acad Sci. 878, 372-387 (1999).</li><li>Kim, J., Yu, W., Kovalski, K., Ossowski, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Requirement+for+specific+proteases+in+cancer+cell+intravasation+as+revealed+by+a+novel+semiquantitative+PCR-based+assay.">Requirement for specific proteases in cancer cell intravasation as revealed by a novel semiquantitative PCR-based assay.</a> Cell. 94, 353-362 (1998).</li><li>Leong, H. S., Steinmetz, N. F., Ablack, A., Destito, G., Zijlstra, A., Stuhlmann, H., Manchester, M., Lewis, 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=Intravital+imaging+of+embryonic+and+tumor+neovasculature+using+viral+nanoparticles.">Intravital imaging of embryonic and tumor neovasculature using viral nanoparticles.</a> Nat Protoc. 5, 1406-1417 (2010).</li><li>Lewis, J. D., Destito, G., Zijlstra, A., Gonzalez, M. J., Quigley, J. P., Manchester, M., Stuhlmann, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viral+nanoparticles+as+tools+for+intravital+vascular+imaging.">Viral nanoparticles as tools for intravital vascular imaging.</a> Nat Med. 12, 354-360 (2006).</li><li>Ossowski, L., Reich, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+malignant+phenotype+of+a+human+carcinoma+conditioned+by+growth+environment.">Changes in malignant phenotype of a human carcinoma conditioned by growth environment.</a> Cell. 33, 323-333 (1983).</li><li>Schmittgen, T. D., Livak, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analyzing+real-time+PCR+data+by+the+comparative+C(T)+method.">Analyzing real-time PCR data by the comparative C(T) method.</a> Nat Protoc. 3, 1101-1108 (2008).</li><li>Testa, J. E., Brooks, P. C., Lin, J. M., Quigley, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eukaryotic+expression+cloning+with+an+antimetastatic+monoclonal+antibody+identifies+a+tetraspanin+(PETA-3/CD151)+as+an+effector+of+human+tumor+cell+migration+and+metastasis.">Eukaryotic expression cloning with an antimetastatic monoclonal antibody identifies a tetraspanin (PETA-3/CD151) as an effector of human tumor cell migration and metastasis.</a> Cancer Res. 59, 3812-3820 (1999).</li><li>Zijlstra, A., Lewis, J., Degryse, B., Stuhlmann, H., Quigley, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Inhibition+of+Tumor+Cell+Intravasation+and+Subsequent+Metastasis+via+Regulation+of+In+Vivo+Tumor+Cell+Motility+by+the+Tetraspanin+CD151.">The Inhibition of Tumor Cell Intravasation and Subsequent Metastasis via Regulation of In Vivo Tumor Cell Motility by the Tetraspanin CD151.</a> Cancer Cell. 13, 221-234 (2008).</li><li>Zijlstra, A., Mellor, R., Panzarella, G., Aimes, R. T., Hooper, J. D., Marchenko, N. D., Quigley, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+quantitative+analysis+of+rate-limiting+steps+in+the+metastatic+cascade+using+human-specific+real-time+polymerase+chain+reaction.">A quantitative analysis of rate-limiting steps in the metastatic cascade using human-specific real-time polymerase chain reaction.</a> Cancer Res. 62, 7083-7092 (2002).</li></ol>

लड़की सीएएम कैंसर जीव विज्ञान और metastatic प्रगति के विभिन्न पहलुओं का अध्ययन करने के लिए एक मॉडल प्रणाली के रूप में दशकों के लिए इस्तेमाल किया गया है. Metastatic क्षमताओं के सिद्धांत मूल्यांकन निद्रा का विश्लेषण...

<table border="1"><tbody><tr><th> अभिकर्मक के नाम </th><th> कंपनी </th><th> सूचीपत्र संख्या </th><th> टिप्पणियां (वैकल्पिक) </th></tr><tr><td> 1X Dulbecco संशोधित ईगल मध्यम (DMEM) </td><td> Invitrogen </td><td> 11995073 </td><td></td></tr><tr><td> Dulbecco फास्फेट buffered खारा (डी पीबीएस) (1X), तरल </td><td> Invitrogen </td><td> 14190250 </td><td> 7.4 पीएच </td></tr><tr><td> Trypsin, EDTA 4Na के साथ 0.05% (1X), तरल </td><td> Invitrogen </td><td> 25300054 </td><td></td></tr><tr><td> खिलाड़ी हैचर </td><td> बेरी हिल </td><td> 1550HA </td><td rowspan="2"> ये इन्क्यूबेटरों सबसे आसानी से उपलब्ध की कुछ कर रहे हैं. हालांकि, विभिन्न उपकरणों वीडियो दिखाएगा. </td></tr><tr><td> खिलाड़ी इनक्यूबेटर </td><td> बेरी हिल </td><td> 1502EA </td><td></td></tr><tr><td> Dremel रोटरी उपकरण </td><td> Dremel </td><td></td><td></td></tr><tr><td> Dremel cutoff पहियों नहीं. 36 </td><td> Dremel </td><td> 409 </td><td></td></tr><tr><td> पोर्टेबल पिपेट सहायता </td><td> मछुआ </td><td> 1368115E </td><td></td></tr><tr><td> कपास Applicators इत्तला दे दी </td><td> मछुआ </td><td> 23-400-001) </td><td></td></tr><tr><td> निषेचित अंडे </td><td> विभिन्न </td><td></td><td> कई अलग अलग विक्रेता इस्तेमाल किया जा सकता है. अंडे लगभग 35-55 ग्राम होना चाहिए. मुर्गियाँ देता है कि उन्हें उम्र के 36 और 55 के बीच सप्ताह होना चाहिए. </td></tr><tr><td> Hemocytometer </td><td> Hausser वैज्ञानिक, VWR </td><td> 15170-090 </td><td></td></tr><tr><td> फाइबर ऑप्टिक माइक्रोस्कोप प्रकाशक </td><td> Amscope </td><td> HL250-AY </td><td> 150W </td></tr><tr><td> 25 गेज सुई </td><td></td><td></td><td></td></tr><tr><td> Sybr ग्रीन निकालें-N-amp ऊतक पीसीआर किट </td><td> सिग्मा Aldrich </td><td> XNATRG </td><td></td></tr></tbody></table>

quantitative analysis of cancer metastasis using an avian embryo model

During metastasis cancer cells disseminate from the primary tumor, invade into surrounding tissues, and spread to distant organs. Metastasis is a complex process that can involve many tissue types, span variable time periods, and often occur deep within organs, making it difficult to investigate and quantify. In addition, the efficacy of the metastatic process is influenced by multiple steps in the metastatic cascade making it difficult to evaluate the contribution of a single aspect of tumor cell behavior. As a consequence, metastasis assays are frequently performed in experimental animals to provide a necessarily realistic context in which to study metastasis. Unfortunately, these models are further complicated by their complex physiology. The chick embryo is a unique in vivo model that overcomes many limitations to studying metastasis, due to the accessibility of the chorioallantoic membrane (CAM), a well-vascularized extra-embryonic tissue located underneath the eggshell that is receptive to the xenografting of tumor cells (figure 1). Moreover, since the chick embryo is naturally immunodeficient, the CAM readily supports the engraftment of both normal and tumor tissues. Most importantly, the avian CAM successfully supports most cancer cell characteristics including growth, invasion, angiogenesis, and remodeling of the microenvironment. This makes the model exceptionally useful for the investigation of the pathways that lead to cancer metastasis and to predict the response of metastatic cancer to new potential therapeutics. The detection of disseminated cells by species-specific Alu PCR makes it possible to quantitatively assess metastasis in organs that are colonized by as few as 25 cells. Using the Human Epidermoid Carcinoma cell line (HEp3) we use this model to analyze spontaneous metastasis of cancer cells to distant organs, including the chick liver and lung. Furthermore, using the Alu-PCR protocol we demonstrate the sensitivity and reproducibility of the assay as a tool to analyze and quantitate intravasation, arrest, extravasation, and colonization as individual elements of metastasis.

Watch this Scientific Journal Video about Quantitative Analysis of Cancer Metastasis using an Avian Embryo Model at JoVE.com

Quantitative Analysis of Cancer Metastasis using an Avian Embryo Model

मात्रात्मक पीसीआर का उपयोग करके, हम प्रदर्शन कैसे अच्छी तरह से स्थापित लड़की सांचा मॉडल मात्रात्मक दूर के अंगों को मानव ट्यूमर कोशिकाओं की मेटास्टेसिस का विश्लेषण के लिए इस्तेमाल किया जा सकता है.

कैंसर मेटास्टेसिस के मात्रात्मक विश्लेषण एक एवियन भ्रूण मॉडल का उपयोग

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

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