旧金山加州大学外科部 雷蒙德沙欣，MD

一，细胞制备<ol><li>大肠癌细胞生长在文化和收获时subconfluent。 </li><li>单细胞悬液，准备在磷酸盐缓冲液放在冰上。 </li></ol>二。肿瘤的制备<ol><li>鼠标与先前成立的皮下大肠肿瘤是安乐死。 </li><li>皮下肿瘤被删除使用无菌技术，并分为2-3毫米件</li><li>肿瘤件保存在磷酸盐缓冲冰生理盐水。 </li></ol>三。鼠标的制备 注：在我们的实验室中，我们使用吸入异氟醚麻醉鼠标，另外，可以使用注射麻醉剂，以达到同样的效果<ol><li>用脚趾捏麻醉深度评估。应该没有退出与脚趾捏反射。 </li><li>在这一点上，可给予抗生素。 </li><li>麻醉的鼠标，这是以前剃光，是正确定位。 </li><li>腹部是prepped用优碘溶液。 </li><li>腹部手术部位的披在无菌的方式。 </li></ol>四。剖腹<ol><li>一个小缺口是在皮肤</li><li>腹壁的肌肉组织是掌握和向上抬起</li><li>进入腹腔和单个刀片的剪刀是用来推动腹内内容</li><li>切口延长至2-3厘米</li></ol>五，暴露盲肠<ol><li>盲肠盲目结束袋标识和exteriorized </li><li>盲肠是孤立的鼠标使用一个预先切开，用无菌纱布休息</li><li>使用温盐水，以保持盲肠湿润</li></ol>六。盲肠壁细胞注射到<ol><li> 27 G或更细的针是用来盲肠壁注入50μL的细胞体积</li><li>取出针</li><li>注射部位的检查，以确保不漏</li><li>盲肠是返回到腹腔</li></ol>七。盲肠肿瘤移植到 注：除了注入盲肠壁细胞，另一种方法是移植到盲肠肿瘤<ol><li>一个8针的数字被放在使用6-0或7-0大中型缝合盲肠</li><li>盲肠壁轻微损坏</li><li>然后，肿瘤定位片</li><li>针绑</li><li>盲肠是返回到腹腔</li></ol>八。鼠标腹壁缝合和恢复<ol><li>鼠标腹壁使用三个使用中断3-0或4-0大中型缝合针封闭</li><li>另外，可以用一个简单的运行针</li><li>在这一点上，可给予手术后止痛药和流体静推</li><li>鼠标是允许从麻醉中恢复。 </li></ol> 注：随着吸入麻醉药，这通常需要30秒九。结果 - 原发灶和肝转移<img src="/files/ftp_upload/old/484_2008_2_11_8/A.png" alt="" /><img src="/files/ftp_upload/old/484_2008_2_11_9/B.png" alt="" /><img src="/files/ftp_upload/old/484_2008_2_11_10/C.png" alt="" /> （三）原发肿瘤 - 原位（A）和新生血管形成的证据（二）; H＆E染色，肿瘤局部侵袭性<img src="/files/ftp_upload/old/484_2008_2_11_11/D.png" alt="" /> 肝转移 - 显示体外（四）

<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> Dig Surg. 22, 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> Comp Med. 50, 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> Int J Cancer. 52, 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> Surg Oncol. 1, 251-256 (1992).</li></ol>

虽然小鼠皮下肿瘤模型很容易的建立和监测，很显然，这种模式无法复制原大肠癌的解剖部位。由于在微环境的差异，皮肤下生长的大肠癌细胞已被证明改变自己的表型和几乎总是失败的进展和转移1,2。事实上，肿瘤对治疗的反应可以相差很大，取决于是否有癌细胞植入宫外孕（皮下）与原位位置3。结直肠癌原位小鼠模型，功能癌细胞生长在其自然位置，高保真复制人类疾病。这两种技术来建?...

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		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
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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>
		</table>
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orthotopic mouse model of colorectal cancer

传统的皮下肿瘤模型不到理想的研究大肠癌。结直肠癌原位小鼠模型，功能癌细胞生长在其自然位置，高保真复制人类疾病。两种技术可用于建立这种模式。这两种技术是相似的，需要曝光的盲肠鼠标麻醉和剖腹探查。的一项技术涉及到盲肠壁注射了大肠癌的细胞悬液。首先是癌症细胞生长的文化，收获时subconfluent，作为一个单细胞悬液准备。缓慢注入的细胞体积小，以避免泄漏。其他技术涉及到盲肠一块皮下肿瘤移植。鼠标与先前成立的皮下大肠肿瘤是安乐死和使用无菌技术的肿瘤被删除。肿瘤一块被划分成小片移植到另一个鼠标。在移植之前，盲肠壁轻微受损，促进肿瘤细胞浸润。发展原发灶和肝转移的时间会有所不同，取决于技术，细胞株，所用的鼠种。这原位小鼠模型为研究大肠癌的自然进程，对大肠癌的测试新的治疗药物的有用。

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

Orthotopic Mouse Model of Colorectal Cancer

两种技术可用于建立这个模型：一个癌症细胞悬液注入到盲肠壁或一块皮下肿瘤移植到盲肠。这种模式是用于研究大肠癌的自然进程和测试新的治疗药物，对大肠癌的。

结直肠癌原位小鼠模型

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

orthotopic-mouse-model-of-colorectal-cancer

Research

JoVE Journal

Biology

44.4K 次观看.  University of California, San Francisco - UCSF. 两种技术可用于建立这个模型：一个癌症细胞悬液注入到盲肠壁或一块皮下肿瘤移植到盲肠。这种模式是用于研究大肠癌的自然进程和测试新的治疗药物，对大肠癌的。