Name: modeling spontaneous metastatic renal cell carcinoma (mrcc) in mice following nephrectomy
Uploaded: 2014-04-29T15:00:00
Description: Watch this Scientific Journal Video about Modeling Spontaneous Metastatic Renal Cell Carcinoma mRCC in Mice Following Nephrectomy at JoVE.com

我们非常感谢博士罗伯特·S·Kerbel（多伦多大学，新宁研究所，加拿大多伦多，加拿大）的技术帮助和专业知识在此过程中发展的实验室。我们还要感谢桑德拉·塞克斯顿博士和罗斯威尔公园癌症研究所部实验动物资源。这项工作是从罗斯威尔公园联盟基金会（以JMLE）裁决的支持。

1。原位肾肿瘤植入<ol><li>细胞培养<ol><li>在此之前原位移植，种植鼠标RENCA LUC细胞单层至75％汇合。 </li><li>以下胰​​蛋白酶消化并重新悬浮于含5％FBS的培养基，离心分离细胞，以1,000 rpm，4℃，5分钟，重复3次，在PBS中洗涤。然后悬浮细胞在无血清培养基，以5×10 4 RENCA LUC / 5μL无血清培养基中的浓度。 注：根据不同的变种或使用细胞系，细胞可以以1:1的比例基?...

<ol>
	<li>Francia, G., Cruz-Munoz, W., Man, S., Xu, P., Kerbel, R. S. <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+advanced+spontaneous+metastasis+for+experimental+therapeutics.">Mouse models of advanced spontaneous metastasis for experimental therapeutics.</a> Nat. Rev. Cancer. 11, 135-141 (2011).</li><li>Souza, B. M., Chaves, K. B., Chammas, R., Schor, N., Bellini, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endostatin+neoadjuvant+gene+therapy+extends+survival+in+an+orthotopic+metastatic+mouse+model+of+renal+cell+carcinoma.">Endostatin neoadjuvant gene therapy extends survival in an orthotopic metastatic mouse model of renal cell carcinoma.</a> Biomed. Pharmacother. 66, 237-241 (2012).</li><li>Amagai, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combination+therapy+of+interleukin-2+and+sorafenib+improves+survival+benefits+and+prevents+spontaneous+pulmonary+metastasis+in+murine+renal+cell+carcinoma+models.">Combination therapy of interleukin-2 and sorafenib improves survival benefits and prevents spontaneous pulmonary metastasis in murine renal cell carcinoma models.</a> Jpn. J. Clin. Oncol. 40, 503-507 (2010).</li><li>Steeg, P. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preclinical+Drug+Development+Must+Consider+the+Impact+on+Metastasis.">Preclinical Drug Development Must Consider the Impact on Metastasis.</a> Clin. Cancer Res. 15, 4529-4530 (2009).</li><li>Guerin, E., Man, S., Xu, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+model+of+postsurgical+advanced+metastatic+breast+cancer+more+accurately+replicates+the+clinical+efficacy+of+antiangiogenic+drugs.">A model of postsurgical advanced metastatic breast cancer more accurately replicates the clinical efficacy of antiangiogenic drugs.</a> Cancer Res. 73, 2743-2748 (2013).</li><li>Day, C. P., Carter, J., Bonomi, C., Hollingshead, M., Merlino, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preclinical+therapeutic+response+of+residual+metastatic+disease+is+distinct+from+its+primary+tumor+of+origin.">Preclinical therapeutic response of residual metastatic disease is distinct from its primary tumor of origin.</a> Int. J. Cancer. 130, 190-199 (2012).</li><li>Ebos, J. M., Kerbel, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antiangiogenic+therapy:+impact+on+invasion,+disease+progression,+and+metastasis.">Antiangiogenic therapy: impact on invasion, disease progression, and metastasis.</a> Nat. Rev. Clin. Oncol. 8, 210-221 (2011).</li><li>Naito, S., Walker, S. M., 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=In+vivo+selection+of+human+renal+cell+carcinoma+cells+with+high+metastatic+potential+in+nude+mice.">In vivo selection of human renal cell carcinoma cells with high metastatic potential in nude mice.</a> Clin. Exp. Metastasis. 7, 381-389 (1989).</li><li>Fidler, I. J., Naito, S., Pathak, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+implantation+in+essential+for+the+selection,+growth+and+metastasis+of+human+renal+cell+cancer+in+nude+mice.">Orthotopic implantation in essential for the selection, growth and metastasis of human renal cell cancer in nude mice.</a> Cancer Metastasis Rev. 9, 145-165 (1990).</li><li>Saiki, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+invasive+and+metastatic+phenotype+in+human+renal+cell+carcinoma.">Characterization of the invasive and metastatic phenotype in human renal cell carcinoma.</a> Clin. Exp. Metastasis. 9, 551-566 (1991).</li><li>Mohanty, S., Xu, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+metastasis+assay.">Experimental metastasis assay.</a> J. Vis. Exp. 42 (1942), (2010).</li><li>Nunez-Cruz, S., Connolly, D. C., Scholler, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+orthotopic+model+of+serous+ovarian+cancer+in+immunocompetent+mice+for+in+vivo+tumor+imaging+and+monitoring+of+tumor+immune+responses.">An orthotopic model of serous ovarian cancer in immunocompetent mice for in vivo tumor imaging and monitoring of tumor immune responses.</a> J. Vis. Exp. (45), (2010).</li><li>Ebos, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerated+metastasis+after+short-term+treatment+with+a+potent+inhibitor+of+tumor+angiogenesis.">Accelerated metastasis after short-term treatment with a potent inhibitor of tumor angiogenesis.</a> Cancer Cell. 15, 232-239 (2009).</li><li>Yang, O. C., Maxwell, P. H., Pollard, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Renal+cell+carcinoma:+translational+aspects+of+metabolism+and+therapeutic+consequences.">Renal cell carcinoma: translational aspects of metabolism and therapeutic consequences.</a> Kidney Int. 84 (4), 667-681 (2013).</li></ol>

此协议的目的是使用同基因肿瘤小鼠模型来描述植入/切除技术来评估临床相关的自发转移性疾病。目前，大多数临床前研究，评估新的实验疗法不包括转移性疾病的研究中，只有少数概括的原发肿瘤生长，手术切除，并最终自发转移扩散的阶段。迄今为止，基因工程小鼠模型（GEMMS）产生的诱发自发性肾细胞疾病进展（见14摘要）不包括后期转移和外科干预策略迄今尚未见报道。因此，自...

死亡的患者的肾细胞癌（RCC）的主要的原因是，通常发生在手术切除原发肿瘤在肾生长的全身性转移疾病。然而，很少有临床前肿瘤模型评价实验治疗的小鼠包括转移性疾病，少仍然忠实地复述局部增长，手术和自发微转移的发生和发展1-3的临床分期。在测试这个差距，已成为新的治疗方法，因为有时出现在动物模型中引人注目的抗肿瘤作用并不总是翻译成同样成功治疗的患者4的评价越来越重要。在结果这种差异可能源于局部的异位或原位原发性肿瘤模型和晚期转移性疾病的5-7之间的差的药物疗效。在碾压的情况下，只有少数研究采用建立了nimal协议，其中包括自发复发的疾病，谁通常有荷瘤肾脏模仿患者全部或部分除去2,3。在小鼠模型中测试这种情况的原因缺乏变化。首先，有高动物成本和肿瘤细胞的选择和转移潜能的固有可变性。例如，人肾细胞系往往很少转移，并且必须在多个回合的主原位植入和转移性选择派生的变种，始终传播并形成远处病灶进行选择（见这样的人细胞系的推导8-10的说明） 。相反，小鼠细胞免疫功能的模型往往表现积极，低细胞数量必须与基质胶，以减少直接的全身播散3注入。其次，在执行适当的植入，手术切除（切除术），和跟踪技术困难（和量化）自发metastati℃生长可以是具有挑战性和几个关键变量采用这种技术时（见讨论的细节）需要考虑。本协议的目的是描述原位移植，切除（切除术），并监测自发转移性肾细胞癌的疾病的基本步骤（以及潜在的缺陷），并提供一个指引，规范（以及更广泛的）科学实验室中使用该评估实​​验治疗的疗效。

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			<td height="21" style="height:21px;width:397px;">DMEM-high glucose with Pyr. And L-Glutamine</td>
			<td style="width:147px;">Corning</td>
			<td style="width:119px;">10-013-CV</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">FBS</td>
			<td style="width:147px;">Invitrogen</td>
			<td style="width:119px;">10437-028</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">0.25% Trypsin EDTA</td>
			<td style="width:147px;">Corning</td>
			<td style="width:119px;">25-053-CL</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">1x DPBS without Calcium &#38; Magnesium</td>
			<td style="width:147px;">Corning</td>
			<td style="width:119px;">21-031-CV</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">Matrigel</td>
			<td style="width:147px;">BD Biosciences</td>
			<td style="width:119px;">354234</td>
			<td>must be kept on ice</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">Artifical tears-lubricant opthalmic ointment</td>
			<td style="width:147px;">Akorn Animal Health</td>
			<td style="width:119px;">17478-162-35</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">Pocket pro pet trimmer</td>
			<td style="width:147px;">Braintree scientific</td>
			<td style="width:119px;">CLP9931B</td>
			<td></td>
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		<tr height="24">
			<td height="24" style="height:24px;width:397px;">Alcohol swab</td>
			<td style="width:147px;">VWR</td>
			<td style="width:119px;">326895</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">Betadine solution swab</td>
			<td style="width:147px;">VWR</td>
			<td style="width:119px;">67618-152-01</td>
			<td></td>
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			<td height="21" style="height:21px;">MICRO DISSECTING sissors straight,blunt - 25mm blades - 4.5&#34;&#160;</td>
			<td style="width:147px;">Southpointe surgical</td>
			<td style="width:119px;">RS-5982</td>
			<td></td>
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			<td height="21" style="height:21px;width:397px;">Iris Forceps, serrated, curved, 10cm long</td>
			<td style="width:147px;">Kent scientific</td>
			<td style="width:119px;">INS15915</td>
			<td>need two of these</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:397px;">10&#181;l Hamilton syringe</td>
			<td style="width:147px;">Hamilton</td>
			<td style="width:119px;">7635-01</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:397px;">30G, 45 degree, RN needle</td>
			<td style="width:147px;">Hamilton</td>
			<td style="width:119px;">7803-07</td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:397px;">Sterile cotton tipped appicator</td>
			<td style="width:147px;">VWR</td>
			<td style="width:119px;">10805-144</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">High temperature cautery kit</td>
			<td style="width:147px;">Kent scientific</td>
			<td style="width:119px;">INS500392</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">5-0 coated Vicryl, conventional cutting needle&#160;</td>
			<td style="width:147px;">Ethicon</td>
			<td style="width:119px;">J834</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">Reflex clip applier for 7mm clips</td>
			<td style="width:147px;">Kent scientific</td>
			<td style="width:119px;">INS500343</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">Reflex clips, 7mm, non-sterile</td>
			<td style="width:147px;">Kent scientific</td>
			<td style="width:119px;">INS500344</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">Removing forceps, 12cm lone</td>
			<td style="width:147px;">Kent scientific</td>
			<td style="width:119px;">INS500347</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">0.9% Sodium Chloride</td>
			<td style="width:147px;">Baxter Healthcare</td>
			<td style="width:119px;">2B1322</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">Buprenorphine 0.01mg/mL</td>
			<td style="width:147px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">25G 5/8&#34; needle</td>
			<td style="width:147px;">VWR</td>
			<td style="width:119px;">BD305122</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">1mL syringe w/out needle</td>
			<td style="width:147px;">VWR</td>
			<td style="width:119px;">BD309659</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:397px;">D-Luciferin</td>
			<td style="width:147px;">Gold Bio technology</td>
			<td style="width:119px;">LUCK-1G</td>
			<td></td>
		</tr>
	</tbody>
</table>

modeling spontaneous metastatic renal cell carcinoma (mrcc) in mice following nephrectomy

其中一个关键的挑战，新的实验治疗肾细胞癌（RCC）提高了测试的模型，忠实地复述早期和晚期转移性疾病进展的发展。典型肿瘤植入模型利用异位或原位原发性肿瘤植入，但很少有全身性的自发转移性疾病，模仿临床上。本协议描述的关键步骤，制定农村信用社疾病进展阶段类似患者。首先，它采用了高转移性小鼠肿瘤细胞系中的同源模型，以显示原位肿瘤细胞植入。方法包括浅表和内部植入与细胞基底膜结合，防止漏电和早期传播子荚膜空间。接着，它描述了程序的荷瘤肾（肾切除术）切除术，具有重要的前和手术后的小鼠护理。最后，它概述了监测和评估的必要步骤微观和宏观转移性疾病的进展，包括生物发光成像，以及提供详细的视觉剖检指南进球全身性疾病的分布。此协议的描述的目的是为了便于普遍使用的临床相关的转移性肾细胞癌模型，以提高未来的治疗试验的预测值。

图1A显示的原理，概述在本协议中总结的详细步骤。有几个重要的因素必须考虑每一步。例如，在步骤1中也示出了两种方法用于子囊肿瘤细胞植入到肾。肿瘤细胞可以被注入到子荚膜空间与一个白色的小泡，确认细胞的渗漏防止通过小心去除针和抽汲过量逸出流体（ 图1B-ⅰ）的局部位置。为了防止泄漏，细胞可植入内部先到达子荚膜空间（ 图1B-ii）中，并?...

Watch this Scientific Journal Video about Modeling Spontaneous Metastatic Renal Cell Carcinoma mRCC in Mice Following Nephrectomy at JoVE.com

Modeling Spontaneous Metastatic Renal Cell Carcinoma mRCC in Mice Following Nephrectomy

可用于临床相关设置评估治疗自发性转移性肾细胞癌（RCC）疾病进展的模型。该协议显示不同的程序原位肾肿瘤细胞植入，适当性肾切除术，最后概述了视觉和生物发光的负担转移和本地化得分尸检指南。

在小鼠模型自发转移性肾细胞癌（MRCC）肾切除

One of the key challenges to improved testing of new experimental therapeutics in renal cell carcinoma (RCC) is the development of models that faithfully ...

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Modeling Spontaneous Metastatic Renal Cell Carcinoma (mRCC) in Mice Following Nephrectomy

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In 2007, Rane presented the first single port nephrectomy for a small non-functioning kidney at the World Congress of Endourology.  Since that time, the ...

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建模在小鼠体内的行程 - 中东与灯丝模型大脑中动脉闭塞

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A preclinical model of invasive bladder cancer was developed in human mucin 1 (MUC1) transgenic (MUC1.Tg) mice for the purpose of evaluating immunotherapy ...

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Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical ...

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The use of murine models to mimic human kidney disease is becoming increasingly common. Our research is focused on the assessment of glomerular function ...