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> Departamento de Cirugía, Universidad de California en San Francisco <br /> Raymond Shaheen, MD </p>

<p class="jove_step"> I. Preparación de la célula </p><ol><li> Las células de cáncer colorrectal en un cultivo y cosecha, cuando subconfluentes. </li><li> Una suspensión de células individuales se prepara en tampón fosfato salino y mantener en hielo. </li></ol><p class="jove_step"> II. Preparación del tumor </p><ol><li> Un ratón con un tumor establecido con anterioridad por vía subcutánea colorrectal es la eutanasia. </li><li> El tumor se elimina por vía subcutánea mediante una técnica estéril y se divide en trozos 2-3 mm </li><l...

<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> Aunque los modelos de ratón de tumores subcutáneos son fáciles de establecer y supervisar, está claro que este modelo no se puede replicar el sitio anatómico original de cáncer colorrectal. Debido a la diferencia en el microambiente de las células de cáncer colorrectal que crecen bajo la piel se ha demostrado que un cambio de fenotipo y casi siempre fallan en el progreso y la metástasis <sup>1,2.</sup> De hecho, la respuesta del tumor a la terapia puede variar enormemente dependiendo de si las células cancerosas se implantan en un...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<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

<p class="jove_content">El modelo tradicional por vía subcutánea tumor es menos que ideal para estudiar el cáncer colorrectal. Ortotópico modelos de ratón de cáncer colorrectal, que cuentan con las células cancerosas que crecen en su ubicación natural, reproducir enfermedades humanas con alta fidelidad. Dos técnicas se pueden utilizar para establecer este modelo. Ambas técnicas son similares y requieren de anestesia del ratón y la laparotomía para la exposición del ciego. Una técnica consiste en la inyección de una suspensión de células cancerosas de colon en la pared del ciego. Las células cancerosas son los primeros en cultivo, cosecha cuando subconfluentes y preparada como una suspensión de células individuales. Un pequeño volumen de las células se inyectaron lentamente para evitar fugas. La otra técnica consiste en el trasplante de un trozo de tumor subcutáneo en el ciego. Un ratón con un tumor establecido con anterioridad por vía subcutánea colorrectal es la eutanasia y el tumor se extirpa mediante una técnica estéril. La pieza del tumor se divide en trozos pequeños para el trasplante a otro ratón. Antes del trasplante, la pared del ciego está ligeramente dañado para facilitar la infiltración de células tumorales. El tiempo necesario para el desarrollo de los tumores primarios y metástasis hepáticas pueden variar dependiendo de la técnica, la línea celular, y las especies de ratón utilizada. Este modelo de ratón ortotópico es útil para estudiar la evolución natural del cáncer colorrectal y ensayo de nuevos agentes terapéuticos contra el cáncer colorrectal.</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">Dos técnicas se pueden utilizar para establecer este modelo: la inyección de una suspensión de células de cáncer en la pared del ciego o el trasplante de un trozo de tumor subcutáneo en el ciego. Este modelo es útil para el estudio de la evolución natural del cáncer colorrectal y ensayo de nuevos agentes terapéuticos contra el cáncer colorrectal.</p>

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