Name: therapy testing in a spheroid-based 3d cell culture model for head and neck squamous cell carcinoma
Uploaded: 2018-04-20T12:30:00
Description: Watch this Scientific Journal Video about Therapy Testing in a Spheroid-based 3D Cell Culture Model for Head and Neck Squamous Cell Carcinoma at JoVE.com

该项目由慕尼黑大学 (FöFoLe 项目-no: 789-781) 拨款资助。

本手稿中所显示的所有研究, 即人类肿瘤标本的使用, 均受《美因茨大学医学》/慕尼黑大学医学中心伦理委员会的事先决定的保护。根据国家法律准则, 病人同意科学使用在其治疗过程中获得的过量生物材料, 因此, 患者已给予知情许可。已按照所有机构、国家和国际人类福利准则进行研究。
1. 头部和见鬼鳞状细胞癌的肿瘤活检
<ol><li>执行一般 (异丙酚和/或七氟醚, 肌肉放松?...

<ol>
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G., 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=The+degree+of+intratumor+mutational+heterogeneity+varies+by+primary+tumor+sub-site.">The degree of intratumor mutational heterogeneity varies by primary tumor sub-site.</a> Oncotarget. 7 (19), 27185-27198 (2016).</li><li>Loyo, 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=Lessons+learned+from+next-generation+sequencing+in+head+and+neck+cancer.">Lessons learned from next-generation sequencing in head and neck cancer.</a> Head Neck. 35 (3), 454-463 (2013).</li><li>Morris, L. G., 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=The+Molecular+Landscape+of+Recurrent+and+Metastatic+Head+and+Neck+Cancers:+Insights+From+a+Precision+Oncology+Sequencing+Platform.">The Molecular Landscape of Recurrent and Metastatic Head and Neck Cancers: Insights From a Precision Oncology Sequencing Platform.</a> JAMA Oncol. , (2016).</li><li>Bauml, J. M., Cohen, R. B., Aggarwal, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunotherapy+for+head+and+neck+cancer:+latest+developments+and+clinical+potential.">Immunotherapy for head and neck cancer: latest developments and clinical potential.</a> Ther Adv Med Oncol. 8 (3), 168-175 (2016).</li><li>Mack, B., 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=Rapid+and+non-enzymatic+in+vitro+retrieval+of+tumour+cells+from+surgical+specimens.">Rapid and non-enzymatic in vitro retrieval of tumour cells from surgical specimens.</a> PLoS One. 8 (1), e55540 (2013).</li><li>Ham, S. L., Joshi, R., Thakuri, P. S., Tavana, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liquid-based+three-dimensional+tumor+models+for+cancer+research+and+drug+discovery.">Liquid-based three-dimensional tumor models for cancer research and drug discovery.</a> Exp Biol Med (Maywood). 241 (9), 939-954 (2016).</li><li>Kelm, J. M., Timmins, N. E., Brown, C. J., Fussenegger, M., Nielsen, L. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Method+for+generation+of+homogeneous+multicellular+tumor+spheroids+applicable+to+a+wide+variety+of+cell+types.">Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types.</a> Biotechnol Bioeng. 83 (2), 173-180 (2003).</li><li>Kunz-Schughart, L. A., Freyer, J. P., Hofstaedter, F., Ebner, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+3-D+cultures+for+high-throughput+screening:+the+multicellular+spheroid+model.">The use of 3-D cultures for high-throughput screening: the multicellular spheroid model.</a> J Biomol Screen. 9 (4), 273-285 (2004).</li><li>Lin, R. Z., Chang, H. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+advances+in+three-dimensional+multicellular+spheroid+culture+for+biomedical+research.">Recent advances in three-dimensional multicellular spheroid culture for biomedical research.</a> Biotechnol J. 3 (9-10), 1172-1184 (2008).</li><li>Weiswald, L. B., Bellet, D., Dangles-Marie, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spherical+cancer+models+in+tumor+biology.">Spherical cancer models in tumor biology.</a> Neoplasia. 17 (1), 1-15 (2015).</li><li>Cancer Genome Atlas, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+genomic+characterization+of+head+and+neck+squamous+cell+carcinomas.">Comprehensive genomic characterization of head and neck squamous cell carcinomas.</a> Nature. 517 (7536), 576-582 (2015).</li><li>Duarte, 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=Isolation+of+head+and+neck+squamous+carcinoma+cancer+stem-like+cells+in+a+syngeneic+mouse+model+and+analysis+of+hypoxia+effect.">Isolation of head and neck squamous carcinoma cancer stem-like cells in a syngeneic mouse model and analysis of hypoxia effect.</a> Oncol Rep. 28 (3), 1057-1062 (2012).</li><li>Reid, P. A., Wilson, P., Li, Y., Marcu, L. G., Bezak, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+understanding+of+cancer+stem+cells:+Review+of+their+radiobiology+and+role+in+head+and+neck+cancers.">Current understanding of cancer stem cells: Review of their radiobiology and role in head and neck cancers.</a> Head Neck. 39 (9), 1920-1932 (2017).</li><li>Cossu, 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=Structural+Insight+into+Inhibitor+of+Apoptosis+Proteins+Recognition+by+a+Potent+Divalent+Smac-Mimetic.">Structural Insight into Inhibitor of Apoptosis Proteins Recognition by a Potent Divalent Smac-Mimetic.</a> PLoS ONE. 7 (11), e49527 (2012).</li><li>Hagemann, J., 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=Spheroid-based+3D+Cell+Cultures+Enable+Personalized+Therapy+Testing+and+Drug+Discovery+in+Head+and+Neck+Cancer.">Spheroid-based 3D Cell Cultures Enable Personalized Therapy Testing and Drug Discovery in Head and Neck Cancer.</a> Anticancer Res. 37 (5), 2201-2210 (2017).</li><li>Worp, H. B., 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=Can+animal+models+of+disease+reliably+inform+human+studies?.">Can animal models of disease reliably inform human studies?.</a> PLoS Med. 7 (3), e1000245 (2010).</li><li>Wilding, J. L., Bodmer, W. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+cell+lines+for+drug+discovery+and+development.">Cancer cell lines for drug discovery and development.</a> Cancer Res. 74 (9), 2377-2384 (2014).</li><li>Chitcholtan, K., Asselin, E., Parent, S., Sykes, P. H., Evans, J. 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我们能够建立一个协议, 以产生可再生的球体从细胞悬浮, 对细胞系和在初步实验中, 原发性人类肿瘤细胞。我们首先评估了两种以前描述的方法, 并确定了 ULA 方法, 这种方法是使用超低粘附表面的培养板, 是一个更安全, 更可靠的三维球体生成。通过结合两种不同的方法来测定读出 (大小/面积和细胞的生存能力), 这种多模式检测是足够敏感, 以确定小的差异组之间。这是迈向技术可行性证明的第一?...

头颈鳞状细胞癌 (HNSCC) 是世界上第六最常见的癌症, 其发病率上升, 粘膜人乳突病毒 (HPV) 感染相关的发病机制, 在大多数情况下, 过量的尼古丁和酒精引起消耗量1,2。虽然较小的肿瘤和前侵入阶段通常是很好的治疗与手术切除, 通常结合宫颈淋巴结清扫, 晚期和复发 HNSCC 治疗仍然是挑战, 由于侵袭性肿瘤侵袭转移扩散和抗辐射和化疗协议3,4,5,6,7,8。最近的研究表明细胞表型的变异很高, 循环和弥散肿瘤细胞的亚型特征刚刚开始9,10。早期对固体、均匀肿瘤质量的信念必须根据最近几年的研究进行修订:11,12,13,14。目前的肿瘤鉴定和关键突变鉴定方法可以识别出一些与治疗耐药性相关的基因, 但仍然是一种成本密集型的方法。而且, 基因型的知识并不一定能对表型及其治疗反应作出可靠的预测。
在改善晚期和复发性疾病的总体和无疾病生存方面, 进展甚少。对于尼古丁-以及与病毒相关的癌症, 目前的治疗方案除了手术外, 还附上了积极的辐射和铂基化学疗法。对 HPV 阴性和阳性癌的反应速率有不同的影响;然而, 这还没有导致一般治疗指南的改变。抗辐射和化疗是在所有肿瘤阶段普遍存在的现象, 并存在于铂基化疗以及靶向治疗 (抗 EGFR;表皮生长因子受体)和最近出现的检查点抑制15。无效的放疗和化疗在吞咽困难、粘膜炎、干口和肾脏或心脏功能下降的风险等方面有很高的病人发病率。预测治疗反应之前, 对每个病人的一般治疗概念的决定似乎是关键的目标, 防止不必要的治疗概念, 副作用和成本。
我们试图建立一个模型, 以测试病人的治疗敏感性目前标准的化疗-辐射, 可纳入常规和质量控制的肿瘤治疗算法从一个技术的站点。远的目标是使用该模型, 而不使用严重改变和老化细胞系, 因为他们不太代表实际的人肿瘤细胞没有他们的变异性和异质性, 因为我们现在知道, 而建立的协议是在不同的细胞系进行。为了独立于商业上可用的细胞线, 我们最近成功地从人类肿瘤标本的原始 HNSCC 细胞中生成了一种称为 "异食癖" 的中间细胞线, 其表面和有限通道上有保守的细胞标记16. 这个异食细胞系应作为一个准备, 以发展的道路上, 然后以后的试验与新鲜的人类癌细胞从肿瘤活检。已经表明, 三维细胞培养细胞在体内的反应不同, 并且比在单分子膜17,18,19,20 中生长的肿瘤药物的治疗更像是癌症药品的管理.21, 主要是由于某些单元格子集22 、23、24的迁移和分异属性的保护。在这里, 我们描述了从中间细胞线和原发性人鳞状细胞癌细胞的球体为基础的三维模型的协议, 以及如何将这种模型整合到头颈外科医生和肿瘤学家的癌症治疗中 (图 1)。

<table>
	<tbody>
		<tr>
			<td>Dulbeccos modified Eagles medium (DMEM)</td>
			<td>Biochrom, Berlin, Germany</td>
			<td>F 0425</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal bovine serum</td>
			<td>Gibco Life Technologies, Paisley, UK</td>
			<td>10500-064</td>
			<td></td>
		</tr>
		<tr>
			<td>penicillin/streptomycin</td>
			<td>Biochrom, Berlin, Germany</td>
			<td>A2212</td>
			<td></td>
		</tr>
		<tr>
			<td>sodium pyruvate</td>
			<td>Biochrom, Berlin, Germany</td>
			<td>L0473</td>
			<td></td>
		</tr>
		<tr>
			<td>non-essential amino acids</td>
			<td>Biochrom, Berlin, Germany</td>
			<td>K0293</td>
			<td></td>
		</tr>
		<tr>
			<td>L-Glutamine</td>
			<td>Biochrom, Berlin, Germany</td>
			<td>K0293</td>
			<td></td>
		</tr>
		<tr>
			<td>Liberase</td>
			<td>Roche Life Sciences, Basel, Switzerland</td>
			<td>5401127001</td>
			<td></td>
		</tr>
		<tr>
			<td>GravityPLUS 3D Culture and Assay Platform</td>
			<td>InSphero, Schlieren, Switzerland</td>
			<td>PB-CS 06-001</td>
			<td></td>
		</tr>
		<tr>
			<td>GravityTRAP plate</td>
			<td>InSphero, Schlieren, Switzerland</td>
			<td>PB-CS-01-001</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultra-low attachment (ULA) culture plates</td>
			<td>Corning, Corning, NY, USA</td>
			<td>4520</td>
			<td></td>
		</tr>
		<tr>
			<td>airway epithelial cell growth medium</td>
			<td>Promocell, Heidelberg, Germany</td>
			<td>C-21060</td>
			<td></td>
		</tr>
		<tr>
			<td>amphotericin B</td>
			<td>Biochrom, Berlin, Germany</td>
			<td>A 2612</td>
			<td></td>
		</tr>
		<tr>
			<td>airway epithelial cell growth medium supplement mix</td>
			<td>Promocell, Heidelberg, Germany</td>
			<td>C39165</td>
			<td></td>
		</tr>
		<tr>
			<td>WST-8 test</td>
			<td>Promocell, Heidelberg, Germany</td>
			<td>PC PK-CA705-CK04</td>
			<td></td>
		</tr>
		<tr>
			<td>Keratinocyte SFMedium + L-Glutamine 500mL</td>
			<td>Invitrogen</td>
			<td>#17005-034</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine Pituitary Extract (BPE), 25mg</td>
			<td>Invitrogen</td>
			<td>#37000015</td>
			<td></td>
		</tr>
		<tr>
			<td>Recombinant human Epithelial Growth Factor 2.5 &#181;g</td>
			<td>Invitrogen</td>
			<td>#37000015</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM High Glucose</td>
			<td>Invitrogen</td>
			<td>#21068-028</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin Streptomycin 10000U/mL Penicillin/ 10000&#181;g/mL Streptomycin</td>
			<td>Invitrogen</td>
			<td>#15140-122</td>
			<td></td>
		</tr>
		<tr>
			<td>F12 Nutrient Mix</td>
			<td>Invitrogen</td>
			<td>#21765-029</td>
			<td></td>
		</tr>
		<tr>
			<td>Glutamax (200 mM L-Alanyl-L-Glutamin-Dipeptide in NaCl)</td>
			<td>Invitrogen</td>
			<td>#35050087</td>
			<td></td>
		</tr>
		<tr>
			<td>HBSS (Ca, Mg)</td>
			<td>Life Technologies</td>
			<td>#14025-092</td>
			<td>(no phenol red)</td>
		</tr>
		<tr>
			<td>1x TrypLE Expres Enzyme</td>
			<td>Invitrogen</td>
			<td>#12604-013</td>
			<td>(no phenol red)</td>
		</tr>
		<tr>
			<td>Accutase (enzymatic cell detachment solution)</td>
			<td>Innovative cell technologies</td>
			<td>Cat# AT104</td>
			<td></td>
		</tr>
		<tr>
			<td>70 &#181;m Falcon cell strainer</td>
			<td>BD Biosciences, USA</td>
			<td>#352350</td>
			<td></td>
		</tr>
	</tbody>
</table>

therapy testing in a spheroid-based 3d cell culture model for head and neck squamous cell carcinoma

目前的治疗方案的晚期和复发头颈部鳞状细胞癌 (HNSCC) 包围辐射和化疗的方法与或没有手术。虽然铂基化疗方案目前代表的是金标准的功效, 并在绝大多数情况下, 新的化疗方案, 即免疫疗法正在出现。然而, 无论是化疗方案的反应率和治疗耐药性机制都难以预测, 而且仍未得到充分了解。目前已知的化疗和耐辐射机制的广泛变化。本研究介绍了一种标准化的高通量体外检测方法, 以评估 HNSCC 细胞系对各种治疗方案的反应, 希望能将个体患者的原细胞作为个性化肿瘤的未来工具。治疗。本试验的目的是将 HNSCC 患者的质量控制标准算法纳入我们的三级护理中心;然而, 这将是未来研究的主题。从实际患者的肿瘤活检中, 技术可行性看起来很有希望。标本然后转移到实验室。活检是机械分离后, 酶消化。细胞, 然后培养在超低黏附细胞培养瓶, 促进可再生, 标准化和自发形成三维球形细胞企业集团。球体然后准备在必要时暴露在化学辐射协议和免疫治疗协议中。最终细胞生存能力和球体大小是治疗敏感性的指标, 因此可以考虑在未来评估患者的可能的治疗反应。这一模式可能是一个宝贵的, 经济高效的工具, 以个性化治疗头颈癌。

我们能够重现性产生球体从单细胞悬浮, 首先从不同的细胞系, 包括专有的异种细胞系, 后来从最初的人类癌细胞的新肿瘤活检中所描述的哈格曼et al.26. 我们评估了两种已建立的球体生成方法;两者是所谓的吊滴 (HD) 方法和超低粘附 (ULA) 方法, 后者是更有效和安全的一个。我们能够突出的优势, ULA 方法相比, HD, 包括更快的球体增长, 更少的问题, 板处?...

Watch this Scientific Journal Video about Therapy Testing in a Spheroid-based 3D Cell Culture Model for Head and Neck Squamous Cell Carcinoma at JoVE.com

Therapy Testing in a Spheroid-based 3D Cell Culture Model for Head and Neck Squamous Cell Carcinoma

我们描述了一个基于球体的三维体外模型的进化, 它使我们能够测试细胞系头颈部鳞状细胞癌实验治疗方案的现行标准, 旨在评估治疗未来人体标本对原代细胞的敏感性和耐药性。

基于球形的头颈部鳞状细胞癌3D 细胞培养模型的治疗试验

Current treatment options for advanced and recurrent head and neck squamous cell carcinoma (HNSCC) enclose radiation and chemo-radiation approaches with ...

The overall goal of this in vitro method is to generate the three dimensional cell culture spheroids that enable the testing of current standard or experimental therapy regimens for head and neck squamous cell carcinoma. This method helps us to understand three dimensional tumor growth of head and neck cancer cells when exposed to radiation or chemo radiation treatment. It remains to be said that handling primary tumor cells is difficult and does not always lead to reliable three dimensional spheroid formation.Demonstrating the procedure will be Sabina Schwenk-Zieger, a technician from our laboratory. While using primary cells from a tumor specimen, place the specimen on a suitable and sterile surface and cut it thoroughly with a sterile, single-use scalpel into very small pieces. After sufficient mechanical separation of the primary tissue, transfer it into a vial containing collagenase 1 and 2 and incubate it for one hour at 37 degrees celsius.Next, sieve the mixture through a 70 micrometer Falcon cell strainer and wash the suspension with HBSS. After successful separation and subsequent washing, transfer the suspension containing one to two million cells into a T75 cell culture flask to grow to subconfluency at 37 degrees celsius for up to ten days. Under a microscope, confirm tumor cell growth.Count the cells in culture. Using an ultra-low adhesion 96-well plate with concave round bottoms, seed 5, 000 primary tumor cells, or 1, 000 to 2, 000 cells of intermediate cell line, in 200 to 300 microliters of media into the wells. Next, culture the cells at 37 degrees celsius.Perform media changes every other day. Pay attention not to aspirate the spheroid with the pipette during media changes. Culture the spheroids until the three dimensional spheroid-shaped cell conglomerates are observed.Keep in mind to exclude the wells with irregular or multiple spheroid formation from further investigation. Subsequently, change the media to media with chemo therapeutics and/or monoclonal antibodies at desired concentrations. Add Cisplatin at the concentrations of 2.5, five, or 10 micromolar or 5-Fluorouracil at 30 micromolar.In this procedure, measure the spheroid size after digital photo documentation on day six, day 10 and day 16 with a graphic software. After centrifugation of the plate at 520g for 2.5 minutes, remove the supernatant. Next, wash the cells with 1x PBS and centrifuge the plate again, followed by removing the supernatant.Next, add 100 microliters of enzymatic cell detachment solution to each vial to allow the spheroids to dissolve. Subsequently, incubate the plate for eight minutes at 37 degrees celsius. Check for successful dissolving of the spheroids under the microscope.Add 100 microliters of DMEM. Next, centrifuge the plate at 520g for 2.5 minutes. Then remove the supernatant and suspend the cells in 100 microliters of DMEM.Subsequently, if necessary, perform a commercially-available colorimetric proliferation assay and read the assay in an ELISA reader. All cell lines could generate reliable spheroids with differences in size and time of readout. Primary cells did form reproducible spheroids in ultra-low attachment plates.The Ki-67 staining reveals the periphery of the culture showing much higher proliferation rates than central cells, mimicking nutrient distribution in solid mucosal tumors that often show necrotic cores. Here, the effect of several chemo therapeutics and radiation on spheroid size. Radiation with two gray prior to treatment with Cisplatin, lead to a significant decrease in spheroid size compared to Cisplatin alone.With further establishment of generating spheroids from primary human cells, this is how the concept of therapy testing could be implemented. Spheroids would be generated after tumor biopsy, and then tested for molecular characteristics and therapy susceptibility. We were able to establish a protocol to generate reproducible spheroids from cell suspensions for both cell lines and primary human tumor cells.This multimodal assay is sufficiently sensitive to identify small differences between groups. In future, the assay could be used to first, assess individual response to standard and experimental therapy protocols. Second, further characterize the tumor cells from fresh specimens.And third, correlate therapy response to molecular patterns. The use of primary cells and the generation of spheroids would allow the preservation of as many characteristics of in vivo tumor growth as possible in combination with a cost-efficient assay for studying individual therapy response.

Research

JoVE Journal

Cancer Research

The Colon-26 Carcinoma Tumor-bearing Mouse as a Model for the Study of Cancer Cachexia

结肠-26癌荷瘤小鼠为癌症恶病质的研究模型

Cancer cachexia is the progressive loss of skeletal muscle mass and adipose tissue, negative nitrogen balance, anorexia, fatigue, inflammation, and ...

科研

癌症研究

A Model for Perineural Invasion in Head and Neck Squamous Cell Carcinoma

在头颈部鳞状细胞癌的神经浸润模型

Perineural invasion (PNI) is found in approximately 40% of head and neck squamous cell carcinomas (HNSCC). Despite multimodal treatment with surgery, ...

A Syngeneic Mouse Model of Metastatic Renal Cell Carcinoma for Quantitative and Longitudinal Assessment of Preclinical Therapies

转移性的型号同源小鼠肾细胞癌的临床前疗法的定量和纵向评估

Renal cell carcinoma (RCC) affects &#62; 60,000 people in the United States annually, and ~ 30% of RCC patients have multiple metastases at the time of ...

A Protocol for Rapid Post-mortem Cell Culture of Diffuse Intrinsic Pontine Glioma (DIPG)

A Protocol for Rapid Post-mortem Cell Culture of Diffuse Intrinsic Pontine Glioma DIPG

一种内在的弥漫性脑胶质瘤脑桥快速验尸细胞培养协议（DIPG）

Diffuse Intrinsic Pontine Glioma (DIPG) is a childhood brainstem tumor that carries a universally fatal prognosis. Because surgical resection is not a ...

Four-color Fluorescence Immunohistochemistry of T-cell Subpopulations in Archival Formalin-fixed, Paraffin-embedded Human Oropharyngeal Squamous Cell Carcinoma Samples

档案福尔马林固定，石蜡包埋的人类口咽鳞状细胞癌样品中T细胞亚群的四色荧光免疫组织化学

The four-color fluorescence immunohistochemistry (IHC) technique is a method to quantify cell populations of interest while taking into account their ...

Fabrication of Tongue Extracellular Matrix and Reconstitution of Tongue Squamous Cell Carcinoma In Vitro

Fabrication of Tongue Extracellular Matrix and Reconstitution of Tongue Squamous Cell Carcinoma In Vitro

舌细胞外基质的制备与舌鳞癌体外重建

In order to construct an effective and realistic model for tongue squamous cell carcinoma (TSCC) in vitro, the methods were created to produce ...

Intramucosal Inoculation of Squamous Cell Carcinoma Cells in Mice for Tumor Immune Profiling and Treatment Response Assessment

小鼠鳞状细胞细胞的肿瘤免疫分析及治疗反应评价

Head and neck squamous cell carcinoma (HNSCC) is a debilitating and deadly disease with a high prevalence of recurrence and treatment failure. To develop ...

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, ...

In Vitro Establishment of a Genetically Engineered Murine Head and Neck Cancer Cell Line using an Adeno-Associated Virus-Cas9 System

使用腺相关病毒-Cas9系统在Vitro建立基因工程的毛因头和颈部癌细胞系

The use of primary normal epithelial cells makes it possible to reproducibly induce genomic alterations required for cellular transformation by ...

Multidimensional Coculture System to Model Lung Squamous Carcinoma Progression

多维共同培养系统模拟肺鳞癌进展

Tumor-stroma interactions play a critical role in the development of lung squamous carcinoma (LUSC). However, understanding how these dynamic interactions ...