Name: orthotopic mouse model of colorectal cancer
Uploaded: 2007-12-04T04:21:00
Description: Watch this Scientific Journal Video about Orthotopic Mouse Model of Colorectal Cancer at JoVE.com

<p class="jove_content">Department of Surgery, University of California, San Francisco <br />Raymond Shaheen, M.D.</p>

<p class="jove_step">I. Cell Preparation</p>
<ol>
<li>Colorectal cancer cells are grown in culture and harvested when subconfluent.</li>
<li>A single cell suspension is prepared in phosphate buffered saline and kept on ice.</li>
</ol>
<p class="jove_step">II. Tumor Preparation</p>
<ol>
<li>A mouse with a previously established subcutaneous colorectal tumor is euthanized. </li>
<li>The subcutaneous tumor is removed using sterile technique and divided into 2-3 mm pieces</li>
<li>The tumor pieces are kept in phosphate buffered saline on ice.</li>
...

<ol>
	<li>Heijstek, M. W., Kraneburg, O., Rinkes Borel, I. H. <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+colorectal+cancer+and+liver+metastases.">Mouse models of colorectal cancer and liver metastases.</a> <em style='font-style: italic'>Dig Surg</em>. <b>22</b>, 16-25 (2005).</li><li>Kobaek-Larsen, M., Thorup, I., Diederichsen, A., Fenger, C., Hoitinga, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review+of+colorectal+cancer+and+its+metastases+in+rodent+models:+comparative+aspects+with+those+in+humans.">Review of colorectal cancer and its metastases in rodent models: comparative aspects with those in humans.</a> <em style='font-style: italic'>Comp Med</em>. <b>50</b>, 16-26 (2000).</li><li>Wilmanns, C., Fan, D., O&apos;Brian, C. A., Bucana, C. D., Fidler, I. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+and+ectopic+organ+environments+differentially+influence+the+sensitivity+of+murine+colon+carcinoma+cells+to+doxorubicin+and+5-fluorouracil.">Orthotopic and ectopic organ environments differentially influence the sensitivity of murine colon carcinoma cells to doxorubicin and 5-fluorouracil.</a> <em style='font-style: italic'>Int J Cancer</em>. <b>52</b>, 98-104 (1992).</li><li>Kashtan, H., Rabau, M., Mullen, J. B., Wong, A. H. C., Roder, J. C., Shiptz, B., Stern, H. S., Gallinger, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intra-rectal+injection+of+tumor+cells:+a+novel+animal+model+of+rectal+cancer.">Intra-rectal injection of tumor cells: a novel animal model of rectal cancer.</a> <em style='font-style: italic'>Surg Oncol</em>. <b>1</b>, 251-256 (1992).</li></ol>

<p class="jove_content">Although mouse subcutaneous tumor models are easy to establish and monitor, it is clear that this model cannot replicate the original anatomic site of colorectal cancer. Due to the difference in microenvironment, colorectal cancer cells growing under the skin have been shown to change their phenotype and almost always fail to progress and metastasize <sup>1,2</sup>. In fact, tumor response to therapy can vary dramatically depending on whether cancer cells are implanted in an ectopic (subcutaneous) versus orthotopic locat...

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		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Cell culture media and components</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>will vary depending on cell line used</td>
</tr>
<tr>
<td>Phosphate Buffered Saline</td>
<td>&nbsp;</td>
<td>BioWhitaker</td>
<td>17-512F</td>
<td>also sold by a number of other vendors</td>
</tr>
<tr>
<td>15 mL centrifuge tubes, polypropylene</td>
<td>&nbsp;</td>
<td>Corning</td>
<td>430790</td>
<td>&nbsp;</td>
<tr>
<td>6 well plates with lid</td>
<td>&nbsp;</td>
<td>Becton Dickinson Labware</td>
<td>353046</td>
<td>&nbsp;</td>
<tr>
<td>Iris forceps, straight, serrated tips</td>
<td>&nbsp;</td>
<td>World Precision Instruments</td>
<td>15914</td>
<td>best to have a pair of these instruments</td>
</tr>
<tr>
<td>Iris dissecting scissors, straight, sharp</td>
<td>&nbsp;</td>
<td>World Precision Instruments</td>
<td>14393</td>
<td>&nbsp;</td>
<tr>
<td>Needle Holder (regular)</td>
<td>&nbsp;</td>
<td>World Precision Instruments</td>
<td>14109</td>
<td>&nbsp;</td>
<tr>
<td>Needle Holder (Castroviejo)</td>
<td>&nbsp;</td>
<td>World Precision Instruments</td>
<td>14137</td>
<td>&nbsp;</td>
<tr>
<td>Cidex Plus</td>
<td>&nbsp;</td>
<td>World Precision Instruments</td>
<td>7364</td>
<td>instruments should be autoclaved periodically, but may also be sterilized by soaking in Cidex</td>
</tr>
<tr>
<td>6-0 or 7-0 suture</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>for cecum; courtesy of Department of Surgery, UCSF</td>
</tr>
<tr>
<td>3-0 or 4-0 suture</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>for abd wall; courtesy of Department of Surgery, UCSF</td>
</tr>
<tr>
<td>Anesthetic machine / Isoflurane</td>
<td>&nbsp;</td>
<td>J.A. Baulch and Associates</td>
<td>3206</td>
<td>also sold by a number of other vendors</td>
</tr>
<tr>
<td>Heating Pad, disposable, 6" x 6"</td>
<td>&nbsp;</td>
<td>Prism Technologies</td>
<td>20419</td>
<td>&nbsp;</td>
<tr>
<td>Tape, orange, 13 mm x 13 m</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>15901F</td>
<td>for restraining anesthetized mouse</td>
</tr>
<tr>
<td>Pro-Cord/Cordless Trimmer</td>
<td>&nbsp;</td>
<td>Oster</td>
<td>78997-010</td>
<td>for shaving mouse fur</td>
</tr>
<tr>
<td>Betadine</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>19-027132</td>
<td>may also be purchased at any medical supply store</td>
</tr>
<tr>
<td>2 x 2 gauze, 3-ply</td>
<td>&nbsp;</td>
<td>Johnson and Johnson</td>
<td>7635</td>
<td>may also be purchased at any medical supply store</td>
</tr>
<tr>
<td>Sterile Field, Barrier, 16" x 29"</td>
<td>&nbsp;</td>
<td>Johnson and Johnson</td>
<td>0905</td>
<td>may also be purchased at any medical supply store</td>
</tr>
<tr>
<td>Cotton tipped applicators, 6"</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>14-960-3Q</td>
<td>may also be purchased at any medical supply store</td>
</tr>
<tr>
<td>Syringe, 10 cc, Luer lock tip</td>
<td>&nbsp;</td>
<td>BD</td>
<td>309604</td>
<td>&nbsp;</td>
<tr>
<td>Syringe, 1 cc, tuberculin, slip tip</td>
<td>&nbsp;</td>
<td>BD</td>
<td>309602</td>
<td>&nbsp;</td>
<tr>
<td>27 1/2 G needle</td>
<td>&nbsp;</td>
<td>BD</td>
<td>305109</td>
<td>&nbsp;</td>
<tr>
<td>30 G needle</td>
<td>&nbsp;</td>
<td>BD</td>
<td>305106</td>
<td>&nbsp;</td>		</tbody>
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orthotopic mouse model of colorectal cancer

The traditional subcutaneous tumor model is less than ideal for studying colorectal cancer. Orthotopic mouse models of colorectal cancer, which feature cancer cells growing in their natural location, replicate human disease with high fidelity. Two techniques can be used to establish this model. Both techniques are similar and require mouse anesthesia and laparotomy for exposure of the cecum. One technique involves injection of a colorectal cancer cell suspension into the cecal wall. Cancer cells are first grown in culture, harvested when subconfluent and prepared as a single cell suspension. A small volume of cells is injected slowly to avoid leakage. The other technique involves transplantation of a piece of subcutaneous tumor onto the cecum. A mouse with a previously established subcutaneous colorectal tumor is euthanized and the tumor is removed using sterile technique. The tumor piece is divided into small pieces for transplantation to another mouse. Prior to transplantation, the cecal wall is lightly damaged to facilitate tumor cell infiltration. The time to developing primary tumors and liver metastases will vary depending on the technique, cell line, and mouse species used. This orthotopic mouse model is useful for studying the natural progression of colorectal cancer and testing new therapeutic agents against colorectal cancer.

Watch this Scientific Journal Video about Orthotopic Mouse Model of Colorectal Cancer at JoVE.com

Orthotopic Mouse Model of Colorectal Cancer

<p class="jove_content">Two techniques can be used to establish this model: injection of a cancer cell suspension into the cecal wall or transplantation of a piece of subcutaneous tumor onto the cecum. This model is useful for studying the natural progression of colorectal cancer and testing new therapeutic agents against colorectal cancer.</p>

The traditional subcutaneous tumor model is less than ideal for studying colorectal cancer. Orthotopic mouse models of colorectal cancer, which feature ...

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