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<article dtd-version="3.0" article-type="Invited Methods Article - JoVE Produced Video">
  <front>
    <journal-meta>
      <journal-id journal-id-type="publisher">jove</journal-id>
      <journal-id journal-id-type="email">editorial-office@jove.com</journal-id>
      <journal-title-group>
        <journal-title>Journal of Visualized Experiments</journal-title>
      </journal-title-group>
      <issn pub-type="ppub">1940-087X</issn>
      <issn pub-type="epub"></issn>
    </journal-meta>
    <article-meta>
      <article-id pub-id-type="publisher-id">JoVE52792</article-id>
      <article-id pub-id-type="manuscript">JoVE52792</article-id>
      <article-categories>
        <subj-group subj-group-type="Article Type">
          <subject>Invited Methods Article - JoVE Produced Video</subject>
        </subj-group>
        <subj-group subj-group-type="Category">
          <subject>JoVE Biology</subject>
        </subj-group>
        <subj-group subj-group-type="Classification">
          <subject>Organoids</subject>
          <subject>Organ Culture Techniques</subject>
          <subject>Organic Chemicals</subject>
        </subj-group>
      </article-categories>
      <title-group>
        <article-title>Chick heart invasion assay for testing the invasiveness of cancer cells and the activity of potentially anti-invasive compounds.</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author" corresp="yes">
          <role content-type="1" />
          <name>
            <surname>Bracke</surname>
            <given-names>Marc</given-names>
            <prefix>Prof..</prefix>
          </name>
          <email>Marc1.Bracke@ugent.be</email>
          <xref ref-type="aff" rid="aff1" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="2" />
          <name>
            <surname>Roman</surname>
            <given-names>Bart</given-names>
          </name>
          <email>bart1.roman@ugent.be</email>
          <xref ref-type="aff" rid="aff2" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="3" />
          <name>
            <surname>Stevens</surname>
            <given-names>Christian</given-names>
          </name>
          <email>chris.stevens@ugent.be</email>
          <xref ref-type="aff" rid="aff3" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="4" />
          <name>
            <surname>Mus</surname>
            <given-names>Liselot</given-names>
          </name>
          <email>liselot.mus@ugent.be</email>
          <xref ref-type="aff" rid="aff4" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="5" />
          <name>
            <surname>Parmar</surname>
            <given-names>Virinder</given-names>
          </name>
          <email>virparmar@gmail.com</email>
          <xref ref-type="aff" rid="aff5" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="6" />
          <name>
            <surname>De Wever</surname>
            <given-names>Olivier</given-names>
          </name>
          <email>olivier.dewever@ugent.be</email>
          <xref ref-type="aff" rid="aff6" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="7" />
          <name>
            <surname>Mareel</surname>
            <given-names>Marc</given-names>
          </name>
          <email>marc.mareel@ugent.be</email>
          <xref ref-type="aff" rid="aff7" />
        </contrib>
        <contrib contrib-type="editor">
          <name>
            <surname>Nguyen</surname>
            <given-names>Nam</given-names>
          </name>
          <degrees>Ph.D.</degrees>
          <email>nam.nguyen@jove.com</email>
          <xref ref-type="aff" rid="aff8" />
        </contrib>
        <aff id="aff1">
          <institution>University of Ghent</institution>
          <institution content-type="position">Laboratory Director</institution>
          <addr-line content-type="addrline1">University Hospital 1P7</addr-line>
          <addr-line content-type="addrline2">De Pintelaan 185</addr-line>
          <country>BELGIUM</country>
          <addr-line content-type="city">Ghent</addr-line>
          <addr-line content-type="state">Flanders</addr-line>
          <addr-line content-type="zipcode">9000</addr-line>
          <phone content-type="primary">3293323007</phone>
        </aff>
        <aff id="aff2">
          <institution>University of Ghent</institution>
          <institution content-type="dept">Department of Sustainable Organic Chemistry and Technology</institution>
          <addr-line content-type="addrline1">Coupure links 653</addr-line>
          <country>BELGIUM</country>
          <addr-line content-type="city">Ghent</addr-line>
          <addr-line content-type="zipcode">9000</addr-line>
          <institution>University of Ghent</institution>
        </aff>
        <aff id="aff3">
          <institution>University of Ghent</institution>
          <institution content-type="dept">Department of Sustainable Organic Chemistry and Technology</institution>
          <addr-line content-type="addrline1">Coupure links 653</addr-line>
          <country>BELGIUM</country>
          <addr-line content-type="city">Ghent</addr-line>
          <addr-line content-type="zipcode">9000</addr-line>
          <institution>University of Ghent</institution>
        </aff>
        <aff id="aff4">
          <institution>University of Ghent</institution>
          <institution content-type="dept">Department of Radiation Oncology and Experimental Cancer Research</institution>
          <addr-line content-type="addrline1">De Pintelaan 185</addr-line>
          <country>BELGIUM</country>
          <addr-line content-type="city">Ghent</addr-line>
          <addr-line content-type="zipcode">9000</addr-line>
          <institution>University of Ghent</institution>
        </aff>
        <aff id="aff5">
          <institution>University of Delhi</institution>
          <institution content-type="dept">Department of Chemistry</institution>
          <addr-line content-type="addrline1">University of Delhi</addr-line>
          <country>INDIA</country>
          <addr-line content-type="city">Delhi</addr-line>
          <addr-line content-type="zipcode">110007</addr-line>
          <institution>University of Delhi</institution>
        </aff>
        <aff id="aff6">
          <institution>University of Ghent</institution>
          <institution content-type="dept">Department of Radiation Oncology and Experimental Cancer Research</institution>
          <addr-line content-type="addrline1">De Pintelaan 185</addr-line>
          <country>BELGIUM</country>
          <addr-line content-type="city">Ghent</addr-line>
          <addr-line content-type="zipcode">9000</addr-line>
          <institution>University of Ghent</institution>
        </aff>
        <aff id="aff7">
          <institution>University of Ghent</institution>
          <institution content-type="dept">Department of Radiation Oncology and Experimental Cancer Research</institution>
          <addr-line content-type="addrline1">De Pintelaan 185</addr-line>
          <country>BELGIUM</country>
          <addr-line content-type="city">Ghent</addr-line>
          <addr-line content-type="zipcode">9000</addr-line>
          <institution>University of Ghent</institution>
        </aff>
        <aff id="aff8">
          <institution>JoVE, Journal of Visualized Experiments</institution>
          <addr-line>1 Alewife Center</addr-line>
          <addr-line>Suite 200</addr-line>
          <addr-line>Cambridge, MA  02140</addr-line>
          <country>UNITED STATES</country>
        </aff>
      </contrib-group>
      <history>
        <date date-type="received">
          <day>03</day>
          <month>10</month>
          <year>2014</year>
        </date>
        <date date-type="rev-recd">
          <day>16</day>
          <month>01</month>
          <year>2015</year>
        </date>
        <date date-type="rev-recd">
          <day>30</day>
          <month>10</month>
          <year>2014</year>
        </date>
        <date date-type="accepted">
          <day>19</day>
          <month>01</month>
          <year>2015</year>
        </date>
      </history>
      <abstract>
        <p>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 between invasive and non-invasive cells, and allows to study the effects of test compounds on tumor invasion. Cancer cells - either as aggregates or single cells - are confronted with fragments of embryonic chick heart. After organ culture in suspension for a few days or weeks the confronting cultures are fixed and embedded in paraffin for histological analysis. The three-dimensional interaction between the cancer cells and the normal tissue is then reconstructed from serial sections stained with hematoxylin-eosin or after immunohistochemical staining for epitopes in the heart tissue or the confronting cancer cells. The assay is consistent with the recent concept that cancer invasion is the result of molecular interactions between the cancer cells and their neighbouring stromal host elements (myofibroblasts, endothelial cells, extracellular matrix components…). In the assay this stromal environment is offered to the cancer cells as a living tissue fragment. Supporting aspects to the relevance of the assay are multiple. Invasion  in the assay is in accordance with the criteria of cancer invasion: progressive occupation and replacement in time and space of the host tissue, and invasiveness or non-invasiveness in vivo of the confronting cells generally correlates with the outcome of the assay. Furthermore, the invasion pattern of cells in vivo, as defined by pathologists, is reflected in the histological images in the assay. Quantitative structure-activity relation (QSAR) analysis of the results obtained with numerous potentially anti-invasive organic congener compounds allowed the study of structure-activity relations for flavonoids and chalcones, and known anti-metastatic drugs used in the clinic (e.g. microtubule inhibitors) inhibit invasion in the assay as well. However, the assay does not take into account immunological contributions to cancer invasion.</p>
      </abstract>
      <kwd-group>
        <kwd>Cancer</kwd>
        <kwd>invasion</kwd>
        <kwd>organ culture</kwd>
        <kwd>histology</kwd>
        <kwd>Heart</kwd>
        <kwd>chicken</kwd>
        <kwd>ecosystem</kwd>
        <kwd>anti-invasive compound</kwd>
        <kwd>polyphenolics</kwd>
        <kwd>structure-activity relation</kwd>
        <kwd>immunohistochemistry.</kwd>
      </kwd-group>
      <funding-group />
      <counts>
        <fig-count count="0" />
      </counts>
      <custom-meta-group>
        <custom-meta>
          <meta-name>Please indicate the &lt;b&gt; full address&lt;/b&gt; at which this article will be &lt;b&gt;filmed&lt;/b&gt;.</meta-name>
          <meta-value>Laboratory of Experimental Cancer Research, University Hospital 1P7, De Pintelaan 185, B-9000 Ghent, Belgium</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Please indicate whether this article will be standard or open access. For information on the differences, &lt;a href="http://www.jove.com/publish/journal-information#7" target="_blank"&gt;click here&lt;/a&gt;</meta-name>
          <meta-value>Standard Access (US$2,400)</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>If the protocol section of this manuscript is longer than 3 pages, are the most important steps highlighted (up to 2.75 pages)?&lt;/br&gt;
&lt;/br&gt;
This is &lt;b&gt;required &lt;/b&gt; for protocols longer than 3 pages, properly formatted (12 pt font, single spaced, space between each step, 1" margins, no indentations). Please see &lt;a href="http://www.jove.com/files/Instructions_for_Authors.pdf"&gt;JoVE Manuscript Instructions for Authors&lt;/a&gt; for more clarification. Remember that the non-highlighted protocol steps will remain in the manuscript and therefore will still be available to the reader. 
&lt;br/&gt;&lt;br/&gt;&lt;b&gt;Type 'Yes' or if needed, provide further explanation.&lt;/b&gt;</meta-name>
          <meta-value>Yes</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Is all of the written text in your manuscript original (not plagiarized, or self-plagiarized)?  
&lt;br&gt;&lt;br&gt;
&lt;em&gt;Exception-&lt;/em&gt; figure legends- if you are using previously published figures you must have permission to re-use the figure from the previous publisher and you must cite the original work in the figure legend.
&lt;br&gt;&lt;br&gt;
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          <meta-value>Yes</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Black and White Image Count</meta-name>
          <meta-value>0</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Color Image Count</meta-name>
          <meta-value>0</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Production Notes</meta-name>
          <meta-value>Request CTM</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Acquisitions Editor</meta-name>
          <meta-value>Nandita Singh</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Assistant who entered lead</meta-name>
          <meta-value>Jennifer Carroll</meta-value>
        </custom-meta>
      </custom-meta-group>
    </article-meta>
  </front>
  <body>
    <fig>
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    <fig>
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    <table-wrap>
      <label>Excel Spreadsheet- Table of Materials/Equipment</label>
      <graphic xlink:href="Kopie van Kopie van JoVE_Materials - final.xlsx" xmlns:xlink="http://www.w3.org/1999/xlink" />
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    <fig>
      <label>Figure 1</label>
      <graphic xlink:href="fig1.docx" xmlns:xlink="http://www.w3.org/1999/xlink" />
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    <fig>
      <label>Figure 2</label>
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    <fig>
      <label>Figure 3</label>
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    <fig>
      <label>Figure 4</label>
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    <fig>
      <label>Figure 5</label>
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    <fig>
      <label>Figure 6</label>
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  </body>
</article>