Name: using the chicken chorioallantoic membrane in vivo model to study gynecological and urological cancers
Uploaded: 2020-01-28T14:00:00
Description: Watch this Scientific Journal Video about Using the Chicken Chorioallantoic Membrane In Vivo Model to Study Gynecological and Urological Cancers at JoVE.com

作者希望感谢Tamanoi博士和Binh Vu博士对这种方法的初步培训。与Eva Koziolek博士的讨论有助于优化这种方法，并对此非常赞赏。如果没有以下来源的资助，这项工作就不可能进行：烟草相关疾病研究计划博士后研究金（27FT-0023， 到ACS，国防部（国防部）卵巢癌研究计划（W81XWH-17-1-0160），NCI/NIH（1R21CA216770），烟草相关疾病研究计划高影响试点奖（27IR-0016）和加州大学洛杉矶分校机构支持， 包括JCCC种子赠款（NCI/NIH P30CA016042）和从研究部副部长办公室到LW的3R补助金。

此处介绍的所有实验都经过加州大学洛杉矶分校 （UCLA） 相关道德委员会的审查和批准。UCLA 机构审查委员会已批准使用非识别的初级人类肿瘤（协议编号 17-000037、17-001169 和 11-001363）。在加州大学洛杉矶分校，动物研究委员会审查不需要使用鸡胚胎的实验;仅当卵子孵化时，才需要协议批准。然而，最佳实践，如AVMA动物安乐死指南，被用来处理鸡胚胎的道德和尽可能避免疼痛。在使用 CAM 模型进?...

<ol>
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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+chicken+chorioallantoic+membrane+model+in+biology,+medicine+and+bioengineering.">The chicken chorioallantoic membrane model in biology, medicine and bioengineering.</a> Angiogenesis. 17 (4), 779-804 (2014).</li><li>Lokman, N. A., Elder, A. S., Ricciardelli, C., Oehler, M. 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F., 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=In+vivo+Wnt+pathway+inhibition+of+human+squamous+cell+carcinoma+growth+and+metastasis+in+the+chick+chorioallantoic+model.">In vivo Wnt pathway inhibition of human squamous cell carcinoma growth and metastasis in the chick chorioallantoic model.</a> Journal of Otolaryngology - Head &amp; Neck Surgery. 45 (1), 26 (2016).</li><li>Canale, 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=Interleukin-27+inhibits+pediatric+B-acute+lymphoblastic+leukemia+cell+spreading+in+a+preclinical+model.">Interleukin-27 inhibits pediatric B-acute lymphoblastic leukemia cell spreading in a preclinical model.</a> Leukemia. 25, 1815 (2011).</li><li>Loos, C., 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=Amino-functionalized+nanoparticles+as+inhibitors+of+mTOR+and+inducers+of+cell+cycle+arrest+in+leukemia+cells.">Amino-functionalized nanoparticles as inhibitors of mTOR and inducers of cell cycle arrest in leukemia cells.</a> Biomaterials. 35 (6), 1944-1953 (2014).</li><li>Rovithi, 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=Development+of+bioluminescent+chick+chorioallantoic+membrane+(CAM)+models+for+primary+pancreatic+cancer+cells:+a+platform+for+drug+testing.">Development of bioluminescent chick chorioallantoic membrane (CAM) models for primary pancreatic cancer cells: a platform for drug testing.</a> Scientific Reports. 7, 44686 (2017).</li><li>Majern&#237;k, 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=Novel+Insights+into+the+Effect+of+Hyperforin+and+Photodynamic+Therapy+with+Hypericin+on+Chosen+Angiogenic+Factors+in+Colorectal+Micro-Tumors+Created+on+Chorioallantoic+Membrane.">Novel Insights into the Effect of Hyperforin and Photodynamic Therapy with Hypericin on Chosen Angiogenic Factors in Colorectal Micro-Tumors Created on Chorioallantoic Membrane.</a> International Journal of Molecular Science. 20 (12), 3004 (2019).</li><li>Swadi, R., 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=Optimising+the+chick+chorioallantoic+membrane+xenograft+model+of+neuroblastoma+for+drug+delivery.">Optimising the chick chorioallantoic membrane xenograft model of neuroblastoma for drug delivery.</a> BMC Cancer. 18 (1), 28 (2018).</li><li>Klingenberg, M., Becker, J., Eberth, S., Kube, D., Wilting, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+chick+chorioallantoic+membrane+as+an+in+vivo+xenograft+model+for+Burkitt+lymphoma.">The chick chorioallantoic membrane as an in vivo xenograft model for Burkitt lymphoma.</a> BMC Cancer. 14 (1), 339 (2014).</li><li>Avram, 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=Standardization+of+A375+human+melanoma+models+on+chicken+embryo+chorioallantoic+membrane+and+Balb/c+nude+mice.">Standardization of A375 human melanoma models on chicken embryo chorioallantoic membrane and Balb/c nude mice.</a> Oncology Reports. 38 (1), 89-99 (2017).</li><li>Zabielska-Koczywas, K., 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=3D+chick+embryo+chorioallantoic+membrane+model+as+an+in+vivo+model+to+study+morphological+and+histopathological+features+of+feline+fibrosarcomas.">3D chick embryo chorioallantoic membrane model as an in vivo model to study morphological and histopathological features of feline fibrosarcomas.</a> BMC Veterinary Research. 13 (1), 201 (2017).</li><li>Skowron, M. 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使用CAM模型的肿瘤扩张和移植允许比体内动物模型更快速和直接地观察到肿瘤生长。此外，一旦完成设备的初始购买，成本就会大大降低，特别是与免疫功能低下小鼠的成本相比。鸡胚胎的初始免疫功能低下状态很容易允许人体和鼠群组织移植。即使有这些优势，CAM 模型也有局限性。如果有必要进行长期治疗研究，短期可能是有益的，也可能有害。CAM模型的免疫功能低下/免疫耐受状态可能使肿?...

老鼠是研究人类疾病（包括恶性肿瘤）的经典模型有机体。作为哺乳动物，它们与人类有许多相似之处。它们的遗传相似性很高，使得转基因操作小鼠基因组提供了对人类疾病基因控制的巨大洞察力。在小鼠的处理和实验方面拥有丰富的经验，因此它们成为生物医学研究的首选模式。然而，除了伦理和科学的关注，关于鼠模型，他们也可以是相当昂贵和耗时2，3。肿瘤的发展可能需要数周甚至数月的时间。仅在一个典型的机构的住房可以运行在数百到数千美元，而肿瘤正在发展。卵巢癌是这个缺点的一个例子，因为它在鼠模型中的生长很容易需要几个月。研究进展的延迟可能会影响卵巢癌患者持续低的5年生存率只有47%（即，在30年内只增加10%的存活率）4。同样，泌尿癌（肾癌、前列腺癌和膀胱癌）占美国所有癌症病例的19%，占癌症相关死亡的11%4。因此，研究妇科和泌尿癌的新型体内方法可以节省实验室大量的时间、人力和资金，即使这种模式只应用于最初的筛选实验。此外，由此带来的研究结果的加速可能对每年被诊断患有这些癌症的177，000人产生重大影响。
鸡CAM模型提供了许多优势，解决上述问题。研究血管生成5，6，肿瘤细胞入侵7，8，和转移7，9，小鸡胚胎CAM模型已经用于研究多种形式的癌症，包括胶质瘤10，11，12，头颈部鳞状细胞癌13，14，白血病15，16，胰腺癌17，结肠直肠癌18。此外，CAM模型已经生成神经母细胞瘤19，伯基特淋巴瘤20，黑色素瘤21，和猫纤维肉瘤22。先前的研究也提出了膀胱癌23和前列腺癌细胞系24的移植，但协议细节有限。鸡蛋不仅比老鼠便宜得多，而且能产生高度可重复的结果25，26。它们显示血管发育迅速，肿瘤移植可在几天内发生，并通过打开的窗口纵向可视化。卵子受精和孵化之间的21天时间，实验可以在几周内完成。此外，成本低、住房需求有限、面积小，很容易进行大型实验，使小鼠研究望而却步。
因此，我们寻求优化CAM模型，用于妇科和泌尿癌的移植。由于早期鸡胚胎27的免疫功能低下，小鼠和人体细胞都可以轻松植入。因此，我们已经成功地移植了卵巢癌、肾癌、前列腺癌和膀胱癌。对于每种肿瘤类型，CAM 欣然接受已建立的小鼠和/或人类肿瘤细胞系。重要的是，新收获的原发性人类肿瘤组织也可以从消化细胞或固体组织碎片中移植，成功率高。每种癌症类型和细胞源都需要优化，我们在这里分享。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>-010 Teflon (PTFE) White 55 Duro Shore D O-Rings</td>
			<td>The O-Ring Store</td>
			<td>TEF010</td>
			<td>Nonstick ring for cell seeding. 1/4&#34;ID X 3/8&#34;OD X 1/16&#34;CS Polytetrafluoroethylene (PTFE).</td>
		</tr>
		<tr>
			<td>C4-2</td>
			<td>ATCC</td>
			<td>CRL-3314</td>
			<td>Human prostate cancer cell line.</td>
		</tr>
		<tr>
			<td>CWR22Rv1</td>
			<td></td>
			<td></td>
			<td>CWR cells were the kind gift of Dr. David Agus (Keck Medicine of University of Southern California)</td>
		</tr>
		<tr>
			<td>Cytokeratin 8/18 Antibody (C-51)</td>
			<td>Novus Biologicals</td>
			<td>NBP2-44929-0.02mg</td>
			<td>Used at a dilution of 1:100 for immunohistochemical analysis of human ovarian CAM tumors.</td>
		</tr>
		<tr>
			<td>D-Luciferin Firefly, potassium salt</td>
			<td>Goldbio</td>
			<td>LUCK-1G</td>
			<td></td>
		</tr>
		<tr>
			<td>Delicate Operating Scissors; Curved; Sharp-Sharp; 30mm Blade Length; 4-3/4 in. Overall Length</td>
			<td>Roboz Surgical</td>
			<td>RS6703</td>
			<td>This is provided as an example. Any similar curved scissors would work as well.</td>
		</tr>
		<tr>
			<td>Dremel 8050-N/18 Micro 8V Max Tool Kit</td>
			<td>Dremel</td>
			<td>8050-N/18</td>
			<td>This kit contains all necessary tools.</td>
		</tr>
		<tr>
			<td>Fertilized chicken eggs (Rhode Island Red - Brown, Lab Grade)</td>
			<td>AA Lab Eggs Inc.</td>
			<td>N/A</td>
			<td>A local egg supplier would need to be identified, as this supplier only delivers regionally.</td>
		</tr>
		<tr>
			<td>HT-1376</td>
			<td>ATCC</td>
			<td>CRL-1472</td>
			<td>Human bladder cancer cell line.</td>
		</tr>
		<tr>
			<td>Hovabator Genesis 1588 Deluxe Egg Incubator Combo Kit</td>
			<td>Incubator Warehouse</td>
			<td>HB1588D-NONE-1102-1588-1357</td>
			<td>Other egg incubators may be used, but their reliability would need to be verified. After implantation, a cell incubator with the CO2 disabled may also be used.</td>
		</tr>
		<tr>
			<td>ID8</td>
			<td></td>
			<td></td>
			<td>Not commercially available, please see PMID: 10753190.</td>
		</tr>
		<tr>
			<td>Incu-Bright Cool Light Egg Candler</td>
			<td>Incubator Warehouse</td>
			<td>1102</td>
			<td>Other candlers may be used; however, this is preferred among those that we have tested. This candler is included in the aforementioned incubator kit.</td>
		</tr>
		<tr>
			<td>Iris Forceps, 10cm, Curved, Serrated, 0.8mm tips</td>
			<td>World Precision Instrument</td>
			<td>15915</td>
			<td>This is provided as an example. Any similar curved forceps would work as well. Multiple brands have been used for this method.</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Clipper Distributing</td>
			<td>0010250</td>
			<td></td>
		</tr>
		<tr>
			<td>IVIS Lumina II In Vivo Imaging System</td>
			<td>Perkin Elmer</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Matrigel Membrane Matrix HC; LDEV-Free</td>
			<td>Corning</td>
			<td>354248</td>
			<td>Extracellular matrix solution</td>
		</tr>
		<tr>
			<td>MyC-CaP</td>
			<td>ATCC</td>
			<td>CRL-3255</td>
			<td>Murine prostate cancer cell line.</td>
		</tr>
		<tr>
			<td>Portable Pipet-Aid XP Pipette Controller</td>
			<td>Drummond Scientific</td>
			<td>4-000-101</td>
			<td>Any similar pipet controller would be appropriate.</td>
		</tr>
		<tr>
			<td>PrecisionGlide Hypodermic Needles</td>
			<td>BD</td>
			<td>305196</td>
			<td>This is provided as an example. Any 18G needle would work similarly.</td>
		</tr>
		<tr>
			<td>RENCA</td>
			<td>ATCC</td>
			<td>CRL-2947</td>
			<td></td>
		</tr>
		<tr>
			<td>Semken Forceps</td>
			<td>Fine Science Tools</td>
			<td>11008-13</td>
			<td>This is provided as an example. Any similar forceps or another style that suits researcher preference would be appropriate.</td>
		</tr>
		<tr>
			<td>SKOV3</td>
			<td>ATCC</td>
			<td>HTB-77</td>
			<td>Human ovarian cancer cell line.</td>
		</tr>
		<tr>
			<td>Specimen forceps</td>
			<td>Electron Microscopy Sciences</td>
			<td>72914</td>
			<td>This is provided as an example. The forceps used for pulling away the shell for bioluminescence imaging are approximately 12.8 cm long with 3 mm-wide tips.</td>
		</tr>
		<tr>
			<td>Sterile Cotton Balls</td>
			<td>Fisherbrand</td>
			<td>22-456-885</td>
			<td>This is provided as an example. Any sterile cotton balls would suffice.</td>
		</tr>
		<tr>
			<td>Stirring Rods with Rubber Policeman; 5mm diameter, 6 in. length</td>
			<td>United Scientific Supplies</td>
			<td>GRPL06</td>
			<td>This is provided as an example. Any similar glass stir rods would work as well.</td>
		</tr>
		<tr>
			<td>T24</td>
			<td>ATCC</td>
			<td>HTB-4</td>
			<td>Human bladder cancer cell line.</td>
		</tr>
		<tr>
			<td>Tegaderm Transparent Dressing Original Frame Style 2 3/8&#34; x 2 3/4&#34;</td>
			<td>Moore Medical</td>
			<td>21272</td>
			<td></td>
		</tr>
		<tr>
			<td>Tissue Culture Dishes, 10 cm diameter</td>
			<td>Corning</td>
			<td>353803</td>
			<td>This is provided as an example. Any similar, sterile 10-cm dish may be used. Tissue culture treatment is not necessary.</td>
		</tr>
		<tr>
			<td>Tygon Clear Laboratory Tubing - 1/4 x 3/8 x 1/16 wall (50 feet)</td>
			<td>Tygon</td>
			<td>AACUN017</td>
			<td>This is provided as an example. Any similarly sized tubing would work as well.</td>
		</tr>
	</tbody>
</table>

using the chicken chorioallantoic membrane in vivo model to study gynecological and urological cancers

小鼠模型是体内癌症研究的基准测试。然而，成本、时间和道德方面的考虑导致了对替代体内癌症模型的呼吁。鸡胆膜 （CAM） 模型提供了一种廉价、快速的替代方案，允许直接可视化肿瘤发育，并适用于体内成像。因此，我们寻求开发一个优化的协议，将妇科和泌尿肿瘤移植到这个模型中，我们在这里介绍。受精后约7天，空气细胞移动到卵子的血管侧，在壳中形成开口。然后，来自鼠和人体细胞系和原发组织的肿瘤可以移植。这些通常播种在细胞外基质和介质的混合物，以避免细胞分散，并提供营养支持，直到细胞招募血管供应。然后，肿瘤在卵子孵化前可能再长14天。通过植入用萤火虫荧光素酶稳稳地转导的细胞，生物发光成像可用于敏感检测膜上的肿瘤生长和整个胚胎的癌细胞。该模型可能用于研究肿瘤原性、入侵、转移和治疗效果。与传统的鼠种模型相比，鸡CAM模型需要的时间和财政资源要少得多。由于卵子具有免疫功能低下和免疫耐受性，任何生物体的组织都有可能被植入，而无需植入人体组织所需的昂贵的转基因动物（如小鼠）。然而，该模型的许多优点也可能有局限性，包括肿瘤生成时间短和免疫功能低下/免疫耐受状态。此外，虽然这里呈现的所有肿瘤类型在鸡胆膜模型中，它们这样做与肿瘤生长的程度不同。

到目前为止，我们发现这种植入方法对于卵巢癌、肾癌、前列腺癌和膀胱癌是成功的。每种产品都经过优化，以确定植入的特定条件，尽管可能具有灵活性。在已测试的肿瘤类型中，卵巢癌的生长远没有那么明显，如果没有生物发光成像的帮助，通常不可见（图1）。然而，在植入区域，用钳子可以感觉到CAM的僵硬。这可能有助于确定在缺乏生物发光...

Watch this Scientific Journal Video about Using the Chicken Chorioallantoic Membrane In Vivo Model to Study Gynecological and Urological Cancers at JoVE.com

Using the Chicken Chorioallantoic Membrane In Vivo Model to Study Gynecological and Urological Cancers

我们提出了鸡胆膜模型作为替代的，可移植的，体内模型，用于妇科和泌尿细胞系和患者衍生肿瘤的移植。

利用活体中鸡胆膜研究妇科和泌尿癌

Mouse models are the benchmark tests for in vivo cancer studies. However, cost, time, and ethical considerations have led to calls for alternative in vivo ...

将肿瘤包到鸡胆膜（CAM）上，可用于肿瘤原性、治疗效果或细胞相声的研究。此方法允许在体内免疫耐症环境中实现具有成本效益的快速肿瘤生长。使用CAM模型，可以使用细胞系甚至患者肿瘤标本快速测试新的治疗方法。该协议最关键的方面是确保适当的 CAM 形成，并且在打开卵子时不会中断 CAM 或血管。在发育的第七天或第八天，当CAM已经完全发育时，关闭卵子旋转器，将大约一到三个鸡蛋放入生物安全柜的蛋架中。关灯后，将一个鸡蛋蜡烛器放在蛋壳上，以识别和标记一个鸡蛋的空气细胞的位置，然后将蛋烛台移动到蛋壳上，以找到一个大血管网络，并尽可能旋转鸡蛋。理想的血管会分支在鸡蛋的中间。使用标记来跟踪用于植入的血管。重新打开发动机罩中的光线后，使用装有碳化硅磨石的无绳旋转工具，直接在空气单元中心上钻一个小孔。当大部分壳体被移除，但白色内膜完好无损时，钻另一个小孔，在那里血管窗口将打开，正如刚刚演示的。当第二个孔准备就绪时，使用 18 测量针轻轻刺穿空气细胞上的白色内膜，并注意白色内膜（而不是 CAM）在整个钻孔区域都被破坏。再次关闭发动机罩后，使用鸡蛋蜡烛器验证空气细胞是否已从蛋端转移到血管上的区域。如有必要，将真空管插入在原气孔周围的移液器控制器中，然后轻轻吸附短脉冲来移动空气单元。标记空气单元在空气 CAM 边界内约 0.5 厘米的新位置的轮廓，并固定一块足够大的包装胶带，以覆盖新空气单元上的孔。在准备另外两个鸡蛋后，将鸡蛋返回到孵化器，并打开任何额外的卵子，在一到三组的实验，如演示。当所有鸡蛋都打开后，将一到三个装有重新定位空气细胞的卵子转移到生物安全柜中的蛋架上，并使用装有圆形切割轮的无绳旋转工具，在空气细胞边界上完全切割一条小线，而不会中断 CAM 或血管。使用弯曲的剪刀，在剩余的空气细胞周围切割，在壳体中创建一个窗口，并验证胚胎的可行性。可行的胚胎将显示广泛的血管，清晰的白细胞素，胚胎运动，和/或可见的心跳。在不可行的胚胎中，CAM可能看起来不透明，很少（如果有任何）存在，胚胎可能很小或没有运动或心跳。使用 Semkin 钳子，从无菌棉球中拉出小块棉花，轻轻印迹可行胚胎的 CAM 表面，以清除任何外壳灰尘和碎屑，然后用四分之一的六 x 七厘米透明薄膜敷料盖住壳开口，无需接触 CAM，即可将卵子返回孵化器，打开的窗户朝上。使用一块蛋架、鸡蛋旋转器的边缘或其他合适的物品来支撑任何不断滚动的鸡蛋。为了准备癌细胞悬浮移植，从感兴趣的癌细胞培养中收获细胞，通过离心沉淀细胞。吸上一液，机械地重新暂停残余介质中的细胞。将细胞悬浮液放在冰上，使用移液器测量培养中的细胞体积。然后以100万至200万细胞的浓度在20至100微升的介质中重新暂停，每毫升细胞外基质和任何生长因子或其他感兴趣的添加剂补充2.7至4毫克。对于癌细胞植入，使用不粘环，将多达六个卵子植入蛋架上，然后将透明薄膜敷料的边缘滚向打开的窗口，以从壳中取出薄膜。检查鸡蛋是否可行和健康。理想的鸡蛋将有一个大容器与较小的分支容器在开放区域的中心。使用弯曲的虹膜钳，将无菌不粘环放在容器上，理想地在分支点上。使用无菌玻璃搅拌杆轻轻擦擦 CAM，将癌细胞悬浮液移液到环的中心。当所有细胞都已交付后，用六厘米五分之一的透明薄膜敷料密封开口，并给卵子贴上适当的植入标记。然后将鸡蛋牢固地放在孵化器中，壳开口朝直。要植入癌细胞，无需使用不粘环，请将细胞悬浮液吸进适当大小的移液器尖端，并按住尖端的顶部小心地弹出移液器尖端，将尖端水平放入无菌的 10 厘米组织培养盘中。装入小菜后，将菜放入37摄氏度的培养箱中15至30分钟，15分钟后检查聚合。少量的液体通常会泄漏出来，因为每个尖端被放置在菜中。使用此液体估计尖端内的聚合程度。当矩阵准备就绪时，将一个尖端放在适当大小的移液器上，然后压压柱塞，以理想地将部分聚合的细胞悬浮液压到 CAM 上，理想地在一个大型发达容器的分支点上。使用这些方法，人类和穆林卵巢癌细胞系和原发性肿瘤片段都成功地移植了与体内其他模型中观察到的形态一致的肿瘤。在测试的肿瘤类型中，卵巢癌的生长不太明显，如果没有生物发光成像的帮助，通常看不到。在植入时添加生长因子或激素的影响也可以使用这些方法进行测试。对于肾癌，从既定细胞系或原发性人类肾肿瘤片段中植入透明细胞肾细胞癌细胞可产生快速、健壮的肿瘤形成。此外，多个人类和穆林前列腺癌细胞系和人类膀胱癌细胞系和原发性肿瘤片已被用来建立肿瘤使用这个模型。必须避免中断 CAM 和血管，并确保 CAM 在打开车窗时不粘附在外壳上。成功移植后，CAM 模型可用于评估肿瘤内不同细胞类型之间感兴趣的治疗或相互作用的疗效。

Research

JoVE Journal

Cancer Research

Coculture Assays to Study Macrophage and Microglia Stimulation of Glioblastoma Invasion

共培养化验，以研究胶质母细胞瘤入侵的巨噬细胞和小胶质细胞刺激

Glioblastoma multiforme (grade IV glioma) is a very aggressive human cancer with a median survival of 1 year post diagnosis. Despite the increased ...

科研

癌症研究

The Drosophila Imaginal Disc Tumor Model: Visualization and Quantification of Gene Expression and Tumor Invasiveness Using Genetic Mosaics

The Drosophila Imaginal Disc Tumor Model: Visualization and Quantification of Gene Expression and Tumor Invasiveness Using Genetic Mosaics

该<em&gt;果蝇</em&gt;成虫盘瘤模型：基因的表达与肿瘤侵袭性的可视化和定量遗传马赛克

Drosophila melanogaster has emerged as a powerful experimental system for functional and mechanistic studies of tumor development and progression in the ...

Live Imaging to Study Microtubule Dynamic Instability in Taxane-resistant Breast Cancers

实时成像来研究抗紫杉烷乳腺癌微管动态失稳

Taxanes such as docetaxel belong to a group of microtubule-targeting agents (MTAs) that are commonly relied upon to treat cancer. However, taxane ...

Detection of Mitochondria Membrane Potential to Study CLIC4 Knockdown-induced HN4 Cell Apoptosis In Vitro

Detection of Mitochondria Membrane Potential to Study CLIC4 Knockdown-induced HN4 Cell Apoptosis In Vitro

线粒体膜电位检测 CLIC4 击倒诱导 HN4 细胞凋亡的实验研究

Depletion of the mitochondrial membrane potential (MMP, &#916;&#936;m) is considered the earliest event in the apoptotic cascade. It even occurs ahead of ...

In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment

In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment

体内在肿瘤微环境中 pH 值、 pO2、氧化还原状态、磷酸盐和谷胱甘肽浓度的 EPR 评价

This protocol demonstrates the capability of low-field electron paramagnetic resonance (EPR)-based techniques in combination with functional paramagnetic ...

Acellular and Cellular Lung Model to Study Tumor Metastasis

脱细胞和细胞肺模型研究肿瘤转移

It is difficult to isolate tumor cells at different points of tumor progression. We created an ex vivo lung model that can show the interaction of tumor ...

Obtaining Cancer Stem Cell Spheres from Gynecological and Breast Cancer Tumors

从妇科和乳腺癌肿瘤获得癌症干细胞球

Cancer stem cells (CSC) are a small population with self-renewal and plasticity which are responsible for tumorigenesis, resistance to treatment and ...

Comparing Metastatic Clear Cell Renal Cell Carcinoma Model Established in Mouse Kidney and on Chicken Chorioallantoic Membrane

比较在小鼠肾脏和鸡胆膜上建立的转移性透明细胞肾细胞癌模型

Metastatic clear cell renal cell carcinoma (ccRCC) is the most common subtype of kidney cancer. Localized ccRCC has a favorable surgical outcome. However, ...

Using Computer-based Image Analysis to Improve Quantification of Lung Metastasis in the 4T1 Breast Cancer Model

利用计算机图像分析提高4T1乳腺癌模型中肺转移的定量

Breast cancer is a devastating malignancy, accounting for 40,000 female deaths and 30% of new female cancer diagnoses in the United States in 2019 alone. ...

Discovery of Metastatic Regulators using a Rapid and Quantitative Intravital Chick Chorioallantoic Membrane Model

使用快速和定量的细胞内胆碱膜模型发现转移调节器

Recent advances in cancer research has illustrated the highly complex nature of cancer metastasis. Multiple genes or genes networks have been found to be ...