Name: assessment of mitochondrial health in cancer-associated fibroblasts isolated from 3d multicellular lung tumor spheroids
Uploaded: 2022-10-21T13:00:00
Description: Watch this Scientific Journal Video about Assessment of Mitochondrial Health in Cancer-Associated Fibroblasts Isolated from 3D Multicellular Lung Tumor Spheroids at JoVE.com

这项工作得到了印度塞尔维亚妇女卓越奖项目（SB/WEA-02/2017）和印度塞尔维亚-早期职业研究奖项目（ECR/2017/000892）的支持。作者LA和SR承认IIT Ropar和MHRD的研究奖学金。MK感谢ICMR的研究奖学金。

1. 细胞培养
<ol><li>在补充有10%FBS和1%青霉素 - 链霉素的RPMI1640培养基中培养人肺腺癌细胞系A549和人单核细胞系THP-1，在5%CO2的加湿室中。</li>
	<li>在补充有10%FBS和1%青霉素 - 链霉素溶液的DMEM培养基中培养MRC-5人肺成纤维细胞，在37°C的加湿室中，在5%CO2。</li>
</ol>2. 使用A549肺腺癌细胞系，MRC5成纤维细胞和THP-1单核细胞制备多细胞肿瘤球状体</p...

<ol>
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本研究介绍了使用改进的悬挂滴法开发包括肿瘤细胞、基质细胞群（即成纤维细胞）和免疫细胞群（即单核细胞）的多细胞肿瘤球体。成纤维细胞和单核细胞/巨噬细胞是构成肿瘤微环境（TME）的最重要群体之一，它们的存在通常与患者预后不良有关16。当存在于TME中时，成纤维细胞发生转化，表现出特定的癌症相关成纤维细胞（CAF）表型，因为肿瘤微环境线索17</sup...

实体瘤由肿瘤微环境（TME）引导的异质细胞群组成，然而，大多数细胞的起源尚未被发现。主要是基质和免疫细胞（成纤维细胞、内皮细胞、单核细胞、巨噬细胞、树突状细胞、B 细胞、T 细胞及其亚群）反映了肺癌、乳腺癌、肾癌和其他实体癌的肿瘤异质性1，2，3。了解每种亚型的起源及其转分化潜力对于开发针对这些癌症的先进疗法至关重要。由于肿瘤类型、部位、分期、样本量限制和患者特异性变异性，在人活检中对这种多样化细胞群的分析本身就面临着一些挑战4。因此，需要一个实验模型，该模型不仅可靠，而且可以模拟体内肿瘤状况，证明其是研究肿瘤-基质串扰及其参与疾病病理生理学的理想选择。
三维（3D）多细胞肿瘤球状体（MCTS）培养物是一种有利的 体外 肿瘤模型系统，因为它们与天然对应物相似。与2D细胞培养模型相比，MCTS可以更好地复制实体瘤的各个方面，包括其空间结构，生理反应，可溶性介质的释放，基因表达模式和耐药机制。此外，MCTS的一个主要优点是它可用于研究肿瘤异质性和肿瘤微环境（TME）。悬挂-下降法是开发和分析 MCTS5 最常用的工具。在这种方法中，带有培养基的不同细胞以液滴的形式悬浮，这允许其以相干的3D聚集体方式生长，并且易于检查。该技术很简单;它不需要很多细胞，并且消除了球状体发育所需的特殊底物（如琼脂糖）6。该方法的另一个优点在于其技术的可重复性。此外，该方法还用于共培养混合细胞群，例如内皮细胞和肿瘤细胞，以模拟早期肿瘤血管生成7。
本研究采用模拟肺肿瘤微环境的悬挂滴法制备了含有肺腺癌细胞、成纤维细胞和单核细胞的多细胞三维肺肿瘤球体。然后分离癌症相关成纤维细胞（CAF）群体以调查线粒体健康状况。开发这些微球背后的主要思想是分离CAF，因为微球中细胞之间的串扰可以将成纤维细胞转化为肌成纤维细胞样激活的CAF状态。其次，这项研究也可能描述异常的ROS产生和线粒体功能障碍如何驱动正常的成纤维细胞走向更具侵略性的CAF表型。结果发现，在肿瘤球体内组装的成纤维细胞具有肌成纤维细胞特征，ROS活性增加，代谢基因表达诱导增加。该协议强调了肿瘤微环境在激活CAF中的重要性，并且可以成为 体外 生成和研究CAF表型特征的优秀模型。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Antibodies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>APC anti-human &#945;-SMA</td>
			<td>R&#38;D systems</td>
			<td>Cat# IC1420A</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-fibroblast microbeads</td>
			<td>Miltenyi Biotec</td>
			<td>Cat# 130-050-601</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell lines</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>A549 lung adenocarcinoma cells</td>
			<td>NCCS Pune</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>MRC-5 fetal lung fibroblasts</td>
			<td>ATCC</td>
			<td>CCL-171</td>
			<td></td>
		</tr>
		<tr>
			<td>THP-1 Human monocytes</td>
			<td>NCCS Pune</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>Chemicals</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>BSA</td>
			<td>Himedia</td>
			<td>Cat# 9048-46-8</td>
			<td></td>
		</tr>
		<tr>
			<td>2,6-dichloroindophenol (DCPIP)</td>
			<td>SRL</td>
			<td>Cat# 55287</td>
			<td></td>
		</tr>
		<tr>
			<td>Calcein-AM</td>
			<td>Thermo Fisher Scientific</td>
			<td>Cat# C3099</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Thermo Fisher Scientific</td>
			<td>Cat# D1306</td>
			<td></td>
		</tr>
		<tr>
			<td>DCFDA</td>
			<td>Sigma</td>
			<td>Cat# D6883</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM</td>
			<td>Gibco</td>
			<td>Cat# 11995073</td>
			<td></td>
		</tr>
		<tr>
			<td>DPBS</td>
			<td>Gibco</td>
			<td>Cat# 14190-144</td>
			<td></td>
		</tr>
		<tr>
			<td>EDTA</td>
			<td>Thermo fisher scientific</td>
			<td>Cat# 17892</td>
			<td></td>
		</tr>
		<tr>
			<td>EGTA</td>
			<td>SRL</td>
			<td>Cat# 62858</td>
			<td></td>
		</tr>
		<tr>
			<td>EZcoun Lactate Dehydrogenase Cell Assay Kit</td>
			<td>HiMedia</td>
			<td>Cat# CCK036</td>
			<td></td>
		</tr>
		<tr>
			<td>FBS</td>
			<td>Gibco</td>
			<td>Cat# 10082147</td>
			<td></td>
		</tr>
		<tr>
			<td>Halt Protease and Phosphatase Inhibitor Cocktail (100X)</td>
			<td>Thermo Fisher Scientific</td>
			<td>Cat# 87786</td>
			<td></td>
		</tr>
		<tr>
			<td>HEPES</td>
			<td>Thermo Fisher Scientific</td>
			<td>Cat# 15630080</td>
			<td></td>
		</tr>
		<tr>
			<td>Horse heart Cytochrome c</td>
			<td>SRL</td>
			<td>Cat# 81551</td>
			<td></td>
		</tr>
		<tr>
			<td>Image-iT Red hypoxia reagent</td>
			<td>Thermo Fisher Scientific</td>
			<td>Cat# H10498</td>
			<td></td>
		</tr>
		<tr>
			<td>JC-1 Dye</td>
			<td>Thermo Fisher Scientific</td>
			<td>Cat# T3168</td>
			<td></td>
		</tr>
		<tr>
			<td>KCl</td>
			<td>Merck</td>
			<td>Cat# P9541</td>
			<td></td>
		</tr>
		<tr>
			<td>MgCl2</td>
			<td>Merck</td>
			<td>Cat# M8266</td>
			<td></td>
		</tr>
		<tr>
			<td>MOPS</td>
			<td>Thermo Fisher Scientific</td>
			<td>Cat# 69824</td>
			<td></td>
		</tr>
		<tr>
			<td>Nacl</td>
			<td>Sigma-Aldrich</td>
			<td>Cat# S9888</td>
			<td></td>
		</tr>
		<tr>
			<td>NADH MB Grade</td>
			<td>SRL</td>
			<td>Cat# 54941</td>
			<td></td>
		</tr>
		<tr>
			<td>NP-40</td>
			<td>Thermo Fisher Scientific</td>
			<td>Cat# 85124</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin/Streptomycin</td>
			<td>Gibco</td>
			<td>Cat# 15140122</td>
			<td></td>
		</tr>
		<tr>
			<td>Phenazine methosulfate (PMS)</td>
			<td>SRL</td>
			<td>Cat# 55782</td>
			<td></td>
		</tr>
		<tr>
			<td>Propidium iodide</td>
			<td>Thermo fisher scientific</td>
			<td>Cat# P1304MP</td>
			<td></td>
		</tr>
		<tr>
			<td>RPMI 1640</td>
			<td>Gibco</td>
			<td>Cat# 11875093</td>
			<td></td>
		</tr>
		<tr>
			<td>Single Cell Lysis Kit</td>
			<td>Thermo Fisher Scientific</td>
			<td>Cat# 4458235</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium ascorbate</td>
			<td>Merck</td>
			<td>Cat# A7631</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium cyanide</td>
			<td>Sigma</td>
			<td>Cat# 205222</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Deoxycholate</td>
			<td>Thermo Fisher Scientific</td>
			<td>Cat# 89904</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium dodecyl sulphate</td>
			<td>Sigma-Aldrich</td>
			<td>Cat# L3771</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium succinate hexahydrate</td>
			<td>SRL</td>
			<td>Cat# 36313</td>
			<td></td>
		</tr>
		<tr>
			<td>Sucrose</td>
			<td>Sigma</td>
			<td>Cat# S0389</td>
			<td></td>
		</tr>
		<tr>
			<td>SuperScript VILO cDNA synthesis kit</td>
			<td>Thermo Fisher Scientific</td>
			<td>Cat# 11754-050</td>
			<td></td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma</td>
			<td>Cat# T8787</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin 0.25% EDTA</td>
			<td>Gibco</td>
			<td>Cat# 25200072</td>
			<td></td>
		</tr>
		<tr>
			<td>Universal SYBR Green Supermix</td>
			<td>BIO-RAD</td>
			<td>Cat# 172-5124</td>
			<td></td>
		</tr>
		<tr>
			<td>Plasticware</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>MACS LS Columns</td>
			<td>Miltenyi Biotec</td>
			<td>Cat# 130-042-401</td>
			<td></td>
		</tr>
		<tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Countess II FL Automated Cell Counter</td>
			<td>Thermo Fisher Scientific</td>
			<td>Cat# AMQAF1000</td>
			<td></td>
		</tr>
		<tr>
			<td>EVOS XL core imaging system</td>
			<td>Thermo Fisher Scientific</td>
			<td>Serial Number F0518-1727-0191</td>
			<td></td>
		</tr>
		<tr>
			<td>LAS X software</td>
			<td>Leica Microsystems</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Leica fluorescent inverted microscope</td>
			<td>s</td>
			<td>DMi8 automated S/N 424150)</td>
			<td></td>
		</tr>
		<tr>
			<td>Midi MACS separator</td>
			<td>Miltenyi Biotec</td>
			<td>Cat# 130-042-302</td>
			<td></td>
		</tr>
	</tbody>
</table>

assessment of mitochondrial health in cancer-associated fibroblasts isolated from 3d multicellular lung tumor spheroids

癌症相关成纤维细胞（CAF）是肿瘤微环境中最丰富的基质细胞之一，可促进肿瘤生长和进展。肿瘤微环境中的复杂性，包括肿瘤分泌组、低度炎症、缺氧和氧化还原失衡，促进了异型相互作用，并允许无活性的常驻成纤维细胞转化为活性 CAF。CAF 在代谢上与正常成纤维细胞 （NF） 不同，因为它们具有更高的糖酵解活性，产生更高水平的活性氧 （ROS），并且过表达乳酸输出剂 MCT-4，导致线粒体通透性过渡孔 （MPTP） 的打开。这里描述了一种方法来分析从多细胞3D肿瘤球体中分离的活化CAF的线粒体健康，该球状体包括人肺腺癌细胞（A549），人单核细胞（THP-1）和人肺成纤维细胞（MRC5）。肿瘤球体以不同的时间间隔分解，并通过磁激活细胞分选分离出CAF。使用JC-1染料评估CAF的线粒体膜电位，通过2'，7'-二氯二氢荧光素二乙酸酯（DCFDA）染色产生ROS以及分离CAF中的酶活性。分析分离CAF的线粒体健康有助于更好地了解反向Warburg效应，也可用于研究CAF线粒体变化的后果，例如代谢通量和相应的肺癌异质性调控机制。因此，本研究提倡了解肿瘤 - 基质相互作用对线粒体健康的影响。它将提供一个平台来检查线粒体特异性候选药物对CAF作为肿瘤微环境中潜在疗法的有效性，从而防止CAF参与肺癌进展。

图1显示了使用三种不同的细胞群-A549（肺腺癌），MRC-5（成纤维细胞）和THP-1（单核细胞）通过悬挂滴法在显微镜下观察到的第7天和第10天多细胞肿瘤球状体的发展。在第7天，球体紧凑而坚硬，直径为260 ± 5.3 μm，在第10天，球体直径为480±7.5μm（图1A上面板，图1B-D）。第7天和第10天的球体是紧密的聚集体，在整个批次...

Watch this Scientific Journal Video about Assessment of Mitochondrial Health in Cancer-Associated Fibroblasts Isolated from 3D Multicellular Lung Tumor Spheroids at JoVE.com

Assessment of Mitochondrial Health in Cancer-Associated Fibroblasts Isolated from 3D Multicellular Lung Tumor Spheroids

用肺腺癌细胞、成纤维细胞和单核细胞制备多细胞3D肿瘤球体，然后从这些球体中分离癌症相关成纤维细胞（CAF）。将分离的CAF与正常成纤维细胞进行比较，通过研究线粒体跨膜电位，活性氧和酶活性来评估线粒体健康状况。

评估从3D多细胞肺肿瘤球体中分离的癌症相关成纤维细胞的线粒体健康

Cancer-associated fibroblasts (CAFs) are among the most abundant stromal cells present in the tumor microenvironment, facilitating tumor growth and ...

该协议强调了肿瘤微环境在激活癌症相关成纤维细胞或CAF方面的重要性。研究表型特征的体外模型可能是极好的。CAF生物学的主要限制在于原发性肿瘤活检样本中CAF的可用性有限。因此，这种技术可用于研究CAF生物学的各个方面。这项研究描述了与CAF激活相关的异常交叉生产和线粒体功能障碍，导致肿瘤预后不良。该方法的应用将有助于评估肿瘤的创伤相互作用，特别是CAF活化和线粒体健康。以探索新型药物靶点。在执行此技术之前，个人必须具有细胞程序的经验。该技术的视觉演示将允许了解将CAF群体与肿瘤球体分离的关键步骤以及研究CAF生物学的下游应用。Leena Arora小姐，Moyna Kalia小姐和Soumyajit Roy夫人，我实验室的助手进行实验。为了开始在DMEM中生长人肺腺癌A549和人肺成纤维细胞MRC-5贴壁细胞，在RPMI中生长人单核细胞THP-1细胞在37摄氏度的T25烧瓶中在含有5%二氧化碳的加湿室中完全生长培养基。三天后，在80%至85%的细胞汇合度下，用一毫升PBS洗涤A549和MRC-5细胞一分钟。接下来，通过将细胞与500微升0.25%胰蛋白酶EDTA溶液在37摄氏度下孵育5分钟来收获细胞。然后，立即加入四毫升完全生长培养基以灭活胰蛋白酶。将细胞悬液收集到15毫升管中，并以125×g沉淀五分钟。除去上清液并将细胞重悬于5毫升完全生长培养基中。为了建立多细胞肿瘤球状体，使用一毫升体积的细胞计数器计数 A549、MRC-5 和 THP-1 细胞数。计数后，以5：4：1的比例制备细胞悬液，并用完整的DMEM将体积补足至一毫升。接下来，将一滴25微升细胞悬浮混合物滴在90毫米培养皿的盖子上。用 10 毫升无菌水填充盘子底部。小心地将盖子倒置在充满水的水合室上，并将培养皿放入细胞培养箱中三天。第四天，在显微镜下监测球体。要获取图像，请打开电源开关。将90毫米培养皿放在载物台上，然后选择10倍放大倍率。调整镜片并检查细胞以分析细胞聚集和增殖。按提供的冻结和保存按钮以捕获图像。成像后，用新鲜培养基小心地从每个液滴中吸出20微升培养基来更换生长培养基。对于活死映像，打开 Ctr 高级，即开关一，然后打开 CPU，等待软件系统启动。启动后，将含有球体的90毫米培养皿放在倒置荧光显微镜的载物台上。然后，通过选择软件中的荧光选项并选择钙黄绿素和德克萨斯红通道的FITC来观察和捕获10倍放大倍率的图像。接下来，选择选项，实时并查看图像。调整荧光强度以进行适当的优化，然后单击安全按钮。在第7天和第10天使用1毫升移液管和15毫升管收集200个肿瘤，每个球状体。然后通过以125×g离心5分钟来沉淀球状体，并小心地吸出上清液。用200微升PBS洗涤球体，再次离心并弃去上清液。加入400微升0.25%胰蛋白酶EDTA溶液用于球状体崩解，并在37摄氏度下孵育10分钟。进行剧烈移液以使球体完全崩解。通过加入一毫升完整的DMEM生长培养基来中和胰蛋白酶。然后离心球状悬浮液并弃去上清液，然后将沉淀重悬于一毫升完整的DMEM培养基中。如图所示计算单元格总数。为了从肿瘤球体中分离癌症相关成纤维细胞或CAF，在80微升冷磁活化细胞分选或MACS缓冲液中将1 x 10重悬至第七个细胞。将20微升抗成纤维细胞微珠加入细胞悬液中，轻轻敲击试管并在室温下孵育30分钟，充分混合。孵育后，用一毫升冷MACS缓冲液洗涤细胞。然后离心并吸出上清液，然后将沉淀重悬于500微升MACS缓冲液中。对于基于磁珠的细胞分离，通过用三毫升MACS缓冲液冲洗MACS柱来制备MACS色谱柱。将细胞悬液放入色谱柱中，并收集含有未标记细胞群的流经。接下来，用三毫升MACS缓冲液清洗色谱柱三次，然后将其从分离器中取出。然后将色谱柱放在收集管上。通过加入五毫升MACS缓冲液并将柱塞牢固地推入柱中来收集抗成纤维细胞微珠标记的细胞。离心标记的细胞，然后再进行下游应用。计数分离的CAF的数量，并使用大约6 x 10至第四个细胞进行流式细胞术分析。用PBS洗涤细胞一次，然后离心并吸出上清液。接下来，加入100微升细胞透化缓冲液，并将细胞在4摄氏度下孵育30分钟，间歇涡旋以维持单细胞悬液。同样，在透化缓冲液中离心并重悬细胞后，加入两微升APC偶联的抗人α SMA抗体，并在4摄氏度下孵育45分钟。孵育后，加入一毫升透化缓冲液并离心以除去多余的抗体。将沉淀重悬于400微升透化缓冲液中，用于流式细胞术。在根据细胞的前向和侧向散射区分细胞群后选择 ACTA2 阳性细胞。选择单重态群体，然后选择在单参数直方图上显示为单个峰的ACTA2阳性细胞。这里显示了多细胞肿瘤球状体的发展和活死体分析。在第七天，球体紧凑而坚硬，直径约为260微米。在第10天，球体的直径为480微米。在细胞活力测定中，观察到碘化丙啶荧光降低，钙黄绿素-AM荧光从第7天到第10天增加。缺氧染色显示第10天球状体中心的缺氧核心。此外，观察到E-钙粘蛋白和Mki-67基因表达随着肿瘤球状体形成天数的增加而上调。从第10天肿瘤球体分离的CAF约为69%ACTA2阳性。与MRC-5成纤维细胞相比，CAF在ACTA2中显示出3倍的上调，在1型α 2链中显示出10倍的上调。与第7天相比，观察到第10天肿瘤球状体的ROS水平显着增加，表明ROS生成可能参与CAF激活。从第7天和第10天肿瘤球体中分离的CAF中观察到细胞ROS水平显着增加。此外，通过JC-1染色观察到线粒体膜电位的改变。与正常成纤维细胞相比，在CAF中观察到GLUT1和MCT4基因表达的诱导。与正常成纤维细胞相比，在CAF中观察到乳酸脱氢酶，细胞色素C氧化酶和琥珀酸脱氢酶活性的显着诱导。A549、MRC-5 和 THP-1 细胞的比例必须为 5：4：1 才能成功开发 3D 多细胞肿瘤球状体。成纤维细胞群对肿瘤球状体的刚性至关重要。与阳离子溶液和表征类似，也可以使用该肿瘤球体来获得肿瘤相关的巨噬细胞群，这是肿瘤进展的帮凶。这种球状体分析将帮助我们了解癌细胞如何参与CAFs激活，也可用于检查线粒体特异性候选药物。

Research

JoVE Journal

Cancer Research

Lung Tumor Cell Recruitment Assay

肺肿瘤细胞招募分析

To investigate the molecular mechanisms governing tumor metastasis, various assays using the mouse as a model animal have been proposed. Here, we ...

科研

癌症研究

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 ...

The Establishment of a Lung Colonization Assay for Circulating Tumor Cell Visualization in Lung Tissues

肺定植法在肺部组织循环肿瘤细胞可视化中的建立

Metastasis is the major cause of cancer death. The role of circulating tumor cells (CTCs) in promoting cancer metastasis, in which lung colonization by ...

A 3D Spheroid Model as a More Physiological System for Cancer-Associated Fibroblasts Differentiation and Invasion In Vitro Studies

3D球形模型作为癌症相关纤维细胞分化和入侵体内研究的更多生理系统

Defining the ideal model for an in vitro study is essential, mainly if studying physiological processes such as differentiation of cells. In the tumor ...

Establishment and Characterization of Small Bowel Neuroendocrine Tumor Spheroids

小肠神经内分泌肿瘤类球体的建立与特征

Small bowel neuroendocrine tumors (SBNETs) are rare cancers originating from enterochromaffin cells of the gut. Research in this field has been limited ...

Detection of Lung Tumor Progression in Mice by Ultrasound Imaging

超声成像对小鼠肺肿瘤进展的检测

With ~1.6 million victims per year, lung cancer contributes tremendously to the worldwide burden of cancer. Lung cancer is partly driven by genetic ...

A Mouse Model to Investigate the Role of Cancer-Associated Fibroblasts in Tumor Growth

研究癌症相关成纤维细胞在肿瘤生长中的作用的小鼠模型

Cancer-associated fibroblasts (CAFs) can play an important role in tumor growth by creating a tumor-promoting microenvironment. Models to study the role ...

Isolation of Primary Cancer-Associated Fibroblasts from a Syngeneic Murine Model of Breast Cancer for the Study of Targeted Nanoparticles

将原发性癌症相关纤维细胞与乳腺癌的合成素 Murine 模型分离，用于靶向纳米粒子的研究

Cancer-associated fibroblasts (CAFs) are key actors in the context of the tumor microenvironment. Despite being reduced in number as compared to tumor ...

Exploring Mitochondrial Energy Metabolism of Single 3D Microtissue Spheroids Using Extracellular Flux Analysis

利用细胞外通量分析探索单个3D微组织球体的线粒体能量代谢

Three-dimensional (3D) cellular aggregates, termed spheroids, have become the forefront of in vitro cell culture in recent years. In contrast to culturing ...

Establishing 3-Dimensional Spheroids from Patient-Derived Tumor Samples and Evaluating their Sensitivity to Drugs

从患者来源的肿瘤样本中建立三维微球并评估其对药物的敏感性

Despite remarkable advances in understanding tumor biology, the vast majority of oncology drug candidates entering clinical trials fail, often due to a ...