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> Department of Surgery, University of California, San Francisco <br /> Raymond Shaheen, MD </p>

<p class="jove_step"> I. Cell Preparation </p><ol><li> Darmkrebs-Zellen in Kultur gezüchtet und geerntet werden, wenn subkonfluenten. </li><li> Eine einzelne Zelle Suspension wird in Phosphat-gepufferter Saline und auf Eis gehalten. </li></ol><p class="jove_step"> II. Tumor Vorbereitung </p><ol><li> Eine Maus mit einem vorher festgelegten subkutane kolorektalen Tumor eingeschläfert. </li><li> Die subkutane Tumor wird entfernt, indem sterile Technik und in 2-3 mm Stücke geteilt </li><li> Der Tumor Stücke sind in Phosphat gehalten Kochsalzlö...

<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> Obwohl Maus subkutanen Tumor-Modellen leicht festzustellen und zu überwachen sind, ist es klar, dass dieses Modell nicht replizieren kann die ursprüngliche anatomische Stelle von Darmkrebs. Aufgrund der Differenz in Mikroumgebung haben kolorektalen Krebszellen wachsen unter der Haut gezeigt, ihren Phänotyp zu ändern und fast immer keine Fortschritte und metastasieren <sup>1,2.</sup> In der Tat können Ansprechen des Tumors auf die Therapie erheblich variieren je nachdem, ob Krebszellen in einer Eileiterschwangerschaft (subkutan) versus ...

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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>
		</table>
		</div>
			<div>
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orthotopic mouse model of colorectal cancer

<p class="jove_content">Die traditionellen subkutanen Tumor-Modell ist weniger als ideal für das Studium des kolorektalen Karzinoms. Orthotopen Mausmodell des kolorektalen Karzinoms, die Krebszellen wachsen in ihrem natürlichen Standort-Funktion, replizieren menschlichen Krankheiten mit hoher Wiedergabetreue. Zwei Techniken können verwendet werden, um dieses Modell zu etablieren. Beide Techniken sind ähnlich und erfordern Maus Anästhesie und Laparotomie für die Exposition des Blinddarms. Eine besteht darin, Injektion eines kolorektalen Karzinoms Zellsuspension in den Blinddarm Wand. Krebszellen werden zuerst in Kultur angebaut, geerntet, wenn subkonfluenten und hergestellt, wie eine einzelne Zelle Suspension. Ein kleines Volumen der Zellen wird langsam, um ein Auslaufen zu vermeiden injiziert. Die andere Technik beinhaltet die Transplantation von einem Stück des subkutanen Tumor auf den Blinddarm. Eine Maus mit einem vorher festgelegten subkutane kolorektalen Tumor eingeschläfert und der Tumor wird entfernt, indem sterile Technik. Der Tumor Stück ist in kleine Stücke für die Transplantation zu einem anderen Maus unterteilt. Vor der Transplantation wird der Blinddarm Wand leicht beschädigt, um Tumor-Zell-Infiltration zu erleichtern. Die Zeit bis zur Entwicklung von Primärtumoren und Lebermetastasen hängt von der Technik-, Zell-Linie, und die Maus Arten verwendet. Diese orthotopen Mausmodell ist nützlich für die Untersuchung der natürlichen Entwicklung von Darmkrebs und Erprobung neuer Therapeutika gegen den Darmkrebs.</p>

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">Zwei Techniken können verwendet werden, um dieses Modell zu etablieren werden: Injektion einer Krebszelle Suspension in den Blinddarm Wand oder Transplantation von einem Stück des subkutanen Tumor auf den Blinddarm. Dieses Modell ist nützlich für die Untersuchung der natürlichen Entwicklung von Darmkrebs und Erprobung neuer Therapeutika gegen den Darmkrebs.</p>

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