Name: a cancer cell spheroid assay to assess invasion in a 3d setting
Uploaded: 2015-11-20T15:00:00.000+00:00
Duration: 5 min 34 s
Description: Watch this Scientific Journal Video about A Cancer Cell Spheroid Assay to Assess Invasion in a 3D Setting at JoVE.com

由美国国立卫生研究院资助P30 CA051008和T32 CA009686（ATR）的支持。

1.球体产生的<ol><li>制备单细胞悬浮液通过分离用的PBS 70％汇合的贴壁〜癌细胞培养物悬滴培养洗涤随后暴露于0.05％的胰蛋白酶-EDTA溶液。 <ol><li>中和与细胞培养基的胰蛋白酶溶液和计数用细胞悬浮液的等分试样的细胞。注意:特定细胞培养基将取决于细胞系进行测试。遵循ATCC建议媒体，其中该细胞培养基是通常的DMEM + 10％FBS。更多信息可在材料表中找到。 </li><li>执行稀释，以允许每细胞培养基的20微升一滴500-1,000细胞的播种。 </li></ol></li><li>收购10cm皿加入5毫升无菌PBS的底部。注:此步骤可以防止蒸发悬滴。成本较低的"细菌学级"菜可以代替细胞培养级的菜肴中使用，因为细胞不会续作用在培养表面。 </li><li>使用多通道移液管到稀释的细胞悬浮液20微升的液滴转移到盖子的内表面上。 <ol><li>吸取40滴（5排8滴）到10 cm培养皿的盖子。 </li></ol></li><li>倒置的盖子，将在培养皿。孵育悬滴培养物在37℃下72小时，以产生球体。 <ol><li>翻转盖的自信，但控制的方式。注意:反相盖子过快或过慢会引起液滴移动。某些细胞系可以形成在48小时或更少的球状体，而其他的可能需要超过72小时，以产生紧凑的聚集体。 </li></ol></li></ol> 2.球体嵌入到3D矩阵<ol><li>嵌入球体之前过夜解冻的生长因子减少的基底膜材料的等分试样于4℃。 <ol><li>可选:图层井ECM提前预防与组织文化前进潜力球体的互动再面。 <ol><li>吸取200微升每孔ECM在24孔板。 </li><li>倾斜板，以确保对ECM覆盖整个井表面。 </li><li>小心地除去多余的ECM用吸管。 </li><li>孵育板直到井干:3小时在室温或过夜，在4℃。 </li></ol></li></ol></li><li>通过倾斜的组织培养皿的盖子，汇集媒体收集的球体。与球体的媒体转移到1.5 ml离心管。 </li><li>允许10分钟的球体，收于离心管的底部。球状​​体应该是肉眼可见的。 </li><li>混合100微升的基底膜材料用100μl冷的（4℃）的I型胶原蛋白在一个单独的预冷管，保持该混合物在4℃下 ，以防止任一凝胶从固化过早。 <ol><li>使用预冷枪头，如果转让的小卷。 </li><li>提供医疗服务时，混合基底膜材料和I型胶原，以防止空气气泡的形成。注: 气泡后可能妨碍成像的协议，如果他们成为镶嵌在凝胶。 </li></ol></li><li>吸从离心管的40微升底部的球状体，并结合基底膜材料/ I型胶原混合物。要小心，以防止形成气泡混合时。注:该解决方案包含100微升基底膜材料，100微升I型胶原，和40微升的球体将创造足够的材料为4个独立的3D文化。这可以放大或缩小。 </li><li>吸取40微升滴粘性的混合物的成孔上的24孔平板的中心。保持板水平，以防止该混合物从运行到井的侧面。注:在24孔板中，一列 - 4口井 - 建议对每个要测试的条件。 </li><li>放置板INTOA 37℃培养箱并留下静置30分钟。三维文化将在此期间聚合。 </li><li>慢慢地淹没在3D培养1毫升细胞培养基。 <ol><li>确保将其添加时入井，以进一步促进凝胶的聚合媒体是温暖的。 </li><li>关键的一步: 轻轻将介质添加到 3D文化。吹打媒体太快入井可导致从组织培养表面上的三维培养物的脱离。 </li></ol></li></ol> 3.监测和分析球体入侵<ol><li>图片入侵从球体到周围的3D矩阵，在时间点由研究者决定。获得使用倒置显微镜以20倍的物镜的照片。注意:理想的时间点会取决于细胞系被测试不同。电镀后不久更具侵入性的细胞系将开始从球体的出口，因此，采取初步的照片S内的几个小时后，电镀。一般来说，拍摄照片，在0小时（接种后），24小时和48小时。入侵通常最后24至48小时后，电镀。 </li><li>利用图像分析软件，如图像J.定量侵袭能力<ol><li>分析侵入从旋转椭圆体的边缘的小区距离。 <ol><li>使用顺子工具，以纪念半径和/或最大侵入距离。 </li><li>点击顶部菜单中的"分析"，然后单击"测量"，显示长度测量。 </li></ol></li><li>分析入侵的球体外面总有创面积。 <ol><li>使用徒手绘制工具来跟踪球体和/或总侵入区域的边界。 </li><li>点击顶部菜单中的"分析"，然后单击"测量"，显示面积测量。 </li></ol></li><li>使用类似的投资回报率经理工具插件创建测量堆栈的图像。 </li></ol>
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The authors declare that they have no competing financial interests.

本研究评估癌细胞系面板的性能在一个球体侵袭测定（ 表1）。一般地，我们发现，球体的形成增强了更多上皮前瞻性的细胞系，其中细胞 - 细胞连接的存在促进了球体状结构的形成。已知已经历上皮间质转变的细胞系，如MDA-MB-231细胞不形成在最有可能的悬滴培养球状体由于它们的减小E-，N-，和P-cadherin表达16。 一些在球体侵袭测定的步骤需要密切遵守的...

经确定癌症细胞运动是预测转移潜能的，因为这样的行为通过基底膜和进入循环系统1促进侵袭。大多数研究蠕动集中于细胞如何表现在二维（2D）的设置，虽然它变得普遍认识到，细胞在三维（3D）矩阵的运动是更具代表性如何这些相同的细胞将实际的表现在体内 2。三维培养系统正越来越多地用于研究的细胞行为，范围从细胞形态，对生长动力学和药物敏感性3。到3D环境的上下文中监测癌细胞侵袭的愿望已经导致了通过悬滴培养法4，接着通过嵌入这些球体成3D胞涉及三维细胞聚集体（球状体）的产生先前建立的技术合成矩阵（ECM）的胶原蛋白和基底膜材料5组成。该方法旨在通过提供可以很容易地利用在各种实验条件下，比较入侵一个简化的方法来改善这些先前建立的技术。 更传统的方式来评估在体外细胞运动是划痕缠绕法和Transwell小实验6。前者测定描绘细胞运动在二维设置，因此独立的多种功能用于体内侵入关键的， 如蛋白酶活性7。 Transwell小测定可以更好的模型细胞入侵时井刀片涂覆有细胞外基质底物，但是，只有一个参数， 即细胞在相对 ​​膜表面的外观被测量，和细胞侵袭的许多细微差别，因此不是很容易观察到。相对于这些技术中，癌细胞球体侵袭测定（图1 &lt;/强&gt;）允许细胞侵入中的设置，不仅生理学相关的实时监控，而且还允许重要细胞系特有的功能而被可视化，如个人与集体细胞迁移8。这种方法也得到了标准的3D文化生长测定的优点。通过悬滴法细胞聚集的产生最初约束细胞运动，从而使细胞将被诱因侵入后该约束被提升。此外，一旦该约束被抬起，细胞出口将进行在一致的方向，然后可以方便地定量。 用于癌症细胞球体测定中最流行的ECM材料是基底膜和I型胶原，其中每个组件具有在影响转移行为重要和独特的作用。基质胶是由Engelbreth-Holm的群小鼠肉瘤细胞产生的蛋白质的分泌的混合物中，并富集basement的膜蛋白诸如层粘连蛋白，巢蛋白和IV型胶原9。出于这个原因基质胶是以下称为"基膜材料。"这些基底膜材料提供了所需的除了该施加的范围内的效果对细胞行为11许多其它蛋白质癌细胞侵袭10期间整合附着力必需配体。相比较而言，I型胶原，通常从肌腱等致密的胶原结构12的酸消化制备，是在作为结缔组织和间质支持组织的身体和器官的一个主要的结构元件更简单的基质材料。它已被证明胶原的物理特性可以调节的一些细胞运动的特征;例如，胶原纤维在肿瘤基质界面的取向允许癌细胞沿着这些原纤维侵入到基质13时，随后迁移。在这里介绍的试验中，两个I型胶原和基底膜材料被用作工具来研究三维癌细胞 - 基质相互作用。 抑制控制侵入可监控的细胞已经嵌入在三维矩阵后途径的或刺激的效果。细胞可以生长在挂滴剂或经转移到三维培养，这取决于一个漫长的处理是否需要调节入侵期间进行预处理。对于更短的处理，建议该药物可以与采集后的球体悬浮液，以及将包围3D培养，以促进适当的药物暴露于细胞的培养基混合。接着，正常或肿瘤相关的基质细胞可以混合与基质材料，以评估其在调节肿瘤细胞侵袭的作用，或以确定如何旁分泌和自分泌信号影响细胞的行为。这个想法是在一项研究显示，其中山坳共培养对癌细胞和内皮细胞在悬滴导致球状体14内的血管网。最后，将ECM成分也可以改变，如癌细胞的侵袭是通过不同的基板15的影响。下面提出的方法将由此提供用于评估在各种条件癌细胞侵袭的框架。一般来说我们发现，并非所有的细胞系将在悬滴创建球状体和上皮前瞻性细胞系通常形成定期球体。

<table><tbody><tr><td>Matrigel Growth Factor Reduced</td><td>Corning</td><td>CB-40234</td><td></td></tr><tr><td>Collagen Type I, Rat Tail, 100 mg</td><td>Millipore</td><td>08-115</td><td></td></tr><tr><td>DMEM</td><td>Life Technologies</td><td>11995-065</td><td></td></tr><tr><td>RPMI 1640 Medium&amp;nbsp;</td><td>Life Technologies</td><td>11875-093</td><td></td></tr><tr><td>PBS</td><td>Life Technologies</td><td>10010-023</td><td></td></tr><tr><td>0.05% Trypsin-EDTA</td><td>Life Technologies</td><td>25300-054</td><td></td></tr><tr><td>Fetal Bovine Serum, Heat Inactivated</td><td>Omega Scientific</td><td>FB-12</td><td></td></tr><tr><td>100 mm TC-treated Dishes</td><td>Corning Incorporated</td><td>430167</td><td></td></tr><tr><td>24-well TC-treated Plates</td><td>NEST Biotechnlology</td><td>702001</td><td></td></tr><tr><td>Olympus IX-71 Inverted Microscope</td><td></td><td></td><td></td></tr><tr><td>Cell lines were maintained in DMEM + 10% FBS, with the expection of BT-474 and LNCaP cells, which were mantained in RPMI + 10% FBS.</td><td></td><td></td><td></td></tr></tbody></table>

a cancer cell spheroid assay to assess invasion in a 3d setting

The invasive nature of cancer cell lines is thought to correlate with their metastatic potential. Most traditional assays, however, do not examine these invasive features in a three-dimensional environment and the resulting data suffer from reduced biological applicability. Here an approach is presented to visualize the invasive ability of cell lines in a physiologically relevant setting. The cancer cell spheroid invasion assay first utilizes gravity to generate spheroids within drops of media that hang from the lid of a cell culture dish. Next, these spheroids are embedded in a 3D matrix consisting of a mixture of basement membrane materials and type I collagen. Cancer cell egression from the spheroids into the surrounding matrix is then monitored over time. The method described here can be modified to examine invasion after coculture of different cell types, inclusion of drugs/inhibitors, or alterations in extracellular matrix (ECM) constituents.

使用球体侵袭测定（图1），肿瘤细胞系的组为他们以形成球状体，以及用于小区出口的在3D基质植入后表现出组成的基底膜材料和I型胶原蛋白的量的能力进行了测试（ 表1）。这些结果表明，并不是每一个细胞系将产生良好的球状体，其中细胞系具有体外上皮形态倾向于产生定期和光滑聚集体。不同的细胞系在测定侵入的能力也是可变的。已经建立的作为是侵入性...

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A Cancer Cell Spheroid Assay to Assess Invasion in a 3D Setting

This method evaluates cancer cell invasion from spheroids into a surrounding 3D matrix. Spheroids are generated via the hanging drop culture method and then embedded in a matrix comprised of basement membrane materials and type I collagen. Invasion out of the spheroids is subsequently monitored.

癌细胞球体检测，以评估入侵的3D设置

The invasive nature of cancer cell lines is thought to correlate with their metastatic potential.  Most traditional assays, however, do not examine these ...

a-cancer-cell-spheroid-assay-to-assess-invasion-in-a-3d-setting

Research

JoVE Journal

Medicine

32.5K 次观看.  Georgetown University. This method evaluates cancer cell invasion from spheroids into a surrounding 3D matrix. Spheroids are generated via the hanging drop culture method and then embedded in a matrix comprised of basement membrane materials and type I collagen. Invasion out of the spheroids is subsequently monitored. 

视频: A Cancer Cell Spheroid Assay to Assess Invasion in a 3D Setting

Spheroid Assay to Measure TGF-&#946;-induced Invasion

TGF-&beta; has opposing roles in breast cancer progression by acting as a tumor suppressor in the initial phase, but stimulating invasion and metastasis at later stage1,2. Moreover, TGF-&beta; is frequently overexpressed in breast cancer and its expression correlates with poor prognosis and metastasis 3,4. The mechanisms by which TGF-&beta; induces invasion are not well understood.
TGF-&beta; elicits its cellular responses via TGF-&beta; type II (T&beta;RII) and type I (T&beta;RI) receptors. Upon TGF-&beta;-induced heteromeric complex formation, T&beta;RII phosphorylates the T&beta;RI. The activated T&beta;RI initiates its intracellular canonical signaling pathway by phosphorylating receptor Smads (R-Smads), i.e. Smad2 and Smad3. These activated R-Smads form heteromeric complexes with Smad4, which accumulate in the nucleus and regulate the transcription of target genes5. In addition to the previously described Smad pathway, receptor activation results in activation of several other non-Smad signaling pathways, for example Mitogen Activated Protein Kinase (MAPK) pathways6.
To study the role of TGF-&beta; in different stages of breast cancer, we made use of the MCF10A cell system. This system consists of spontaneously immortalized MCF10A1 (M1) breast epithelial cells7, the H-RAS transformed M1-derivative MCF10AneoT (M2), which produces premalignant lesions in mice8, and the M2-derivative MCF10CA1a (M4), which was established from M2 xenografts and forms high grade carcinomas with the ability to metastasize to the lung9. This MCF10A series offers the possibility to study the responses of cells with different grades of malignancy that are not biased by a different genetic background.
For the analysis of TGF-&beta;-induced invasion, we generated homotypic MCF10A spheroid cell cultures embedded in a 3D collagen matrix in vitro (Fig 1). Such models closely resemble human tumors in vivo by establishing a gradient of oxygen and nutrients, resulting in active and invasive cells on the outside and quiescent or even necrotic cells in the inside of the spheroid10. Spheroid based assays have also been shown to better recapitulate drug resistance than monolayer cultures11. This MCF10 3D model system allowed us to investigate the impact of TGF-&beta; signaling on the invasive properties of breast cells in different stages of malignancy.

An assay to quantitatively measure Transforming Growth Factor (TGF)-&#x03B2;-induced invasion in 3-dimensional collagen gels is described. This assay takes advantage of the MCF10A series of cell lines, which represent different stages of breast cancer development. This method can be adopted to be used with other cell lines and might be used to investigate other potential activators or inhibitors of invasion.

球体的检测，以测量TGF -β诱导的入侵

TGF-&beta; has opposing roles in breast cancer progression by acting as a tumor suppressor in the initial phase, but stimulating invasion and metastasis ...

科研

医学

Fully Human Tumor-based Matrix in Three-dimensional Spheroid Invasion Assay

Two-dimensional cell culture-based assays are commonly used in in vitro cancer research. However, they lack several basic elements that form the tumor microenvironment. To obtain more reliable in vitro results, several three-dimensional (3D) cell culture assays have been introduced. These assays allow cancer cells to interact with the extracellular matrix. This interaction affects cell behavior, such as proliferation and invasion, as well as cell morphology. Additionally, this interaction could induce or suppress the expression of several pro- and anti-tumorigenic molecules. Spheroid invasion assay was developed to provide a suitable 3D in vitro method to study cancer cell invasion. Currently, animal-derived matrices, such as mouse sarcoma-derived matrix (MSDM) and rat tail type I collagen, are mainly used in the spheroid invasion assays. Taking into consideration the differences between the human tumor microenvironment and animal-derived matrices, a human myoma-derived matrix (HMDM) was developed from benign uterus leiomyoma tissue. It has been shown that HMDM induces migration and invasion of carcinoma cells better than MSDM. This protocol provided a simple, reproducible, and reliable 3D human tumor-based spheroid invasion assay using the HMDM/fibrin matrix. It also includes detailed instructions on imaging and analysis. The spheroids grow in a U-shaped ultra-low attachment plate within the HMDM/fibrin matrix and invade through it. The invasion is daily imaged, measured, and analyzed using ilastik and Fiji ImageJ software. The assay platform was demonstrated using human laryngeal primary and metastatic squamous cell carcinoma cell lines. However, the protocol is suitable also for other solid cancer cell lines.

Tumor microenvironment is an essential part of cancer growth and invasion. To mimic carcinoma progression, a biologically relevant human matrix is needed. This protocol introduces an improvement for the in vitro three-dimensional spheroid invasion assay by applying a human leiomyoma-based matrix. The protocol also introduces a computer-based cell invasion analysis.

三维流状入侵检测中的全人肿瘤基基质

Two-dimensional cell culture-based assays are commonly used in in vitro cancer research. However, they lack several basic elements that form the tumor ...

癌症研究

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

Defining the ideal model for an in vitro study is essential, mainly if studying physiological processes such as differentiation of cells. In the tumor stroma, host fibroblasts are stimulated by cancer cells to differentiate. Thus, they acquire a phenotype that contributes to the tumor microenvironment and supports tumor progression. By using the spheroid model, we have set up such a 3D in vitro model system, in which we analyzed the role of laminin-332 and its receptor integrin &#945;3&#946;1 in this differentiation process. This spheroid model system not only reproduces the tumor microenvironment conditions in a more accurate way, but also is a very versatile model since it allows different downstream studies, such as immunofluorescent staining of both intra- and extracellular markers, as well as deposited extracellular matrix proteins. Moreover, transcriptional analyses by qPCR, flow cytometry and cellular invasion can be studied with this model. Here, we describe a protocol of a spheroid model to assess the role of CAFs&#39; integrin &#945;3&#946;1 and its ectopically deposited ligand, laminin-332, in differentiation and in supporting the invasion of pancreatic cancer cells.

The goal of this protocol is to establish a 3D in vitro model to study the differentiation of cancer-associated fibroblasts (CAFs) in a tumor bulk-like environment, which can be addressed in different analysis systems, such as immunofluorescence, transcriptional analysis and life cell imaging.

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 ...

Monitoring Cancer Cell Invasion and T-Cell Cytotoxicity in 3D Culture

Significant progress has been made in treating cancer with immunotherapy, although a large number of cancers remain resistant to treatment. A limited number of assays allow for direct monitoring and mechanistic insights into the interactions between tumor and immune cells, amongst which, T-cells play a significant role in executing the cytotoxic response of the adaptive immune system to cancer cells. Most assays are based on two-dimensional (2D) co-culture of cells due to the relative ease of use but with limited representation of the invasive growth phenotype, one of the hallmarks of cancer cells. Current three-dimensional (3D) co-culture systems either require special equipment or separate monitoring for invasion of co-cultured cancer cells and interacting T-cells.
Here we describe an approach to simultaneously monitor the invasive behavior in 3D of cancer cell spheroids and T-cell cytotoxicity in co-culture. Spheroid formation is driven by enhanced cell-cell interactions in scaffold-free agarose microwell casts with U-shaped bottoms. Both T-cell co-culture and cancer cell invasion into type I collagen matrix are performed within the microwells of the agarose casts without the need to transfer the cells, thus maintaining an intact 3D co-culture system throughout the assay. The collagen matrix can be separated from the agarose cast, allowing for immunofluorescence (IF) staining and for confocal imaging of cells. Also, cells can be isolated for further growth or subjected to analyses such as for gene expression or fluorescence activated cell sorting (FACS). Finally, the 3D co-culture can be analyzed by immunohistochemistry (IHC) after embedding and sectioning. Possible modifications of the assay include altered compositions of the extracellular matrix (ECM) as well as the inclusion of different stromal or immune cells with the cancer cells.

The presented approach simultaneously evaluates cancer cell invasion in 3D spheroid assays and T-cell cytotoxicity. Spheroids are generated in a scaffold-free agarose multi-microwell cast. Co-culture and embedding in type I collagen matrix are performed within the same device which allows to monitor cancer cell invasion and T-cell mediated cytotoxicity.

监测三D培养中的癌细胞入侵和T细胞毒性

Significant progress has been made in treating cancer with immunotherapy, although a large number of cancers remain resistant to treatment. A limited ...

癌症生物学中的 3D 模型:异球体中的侵袭动力学

Alternative Strategy to Analyze In Vitro Cell Invasion of 3D Cultures

Spheroid culture is a 3D model that provides an improved replication of the in vivo microenvironment compared to traditional two-dimensional (2D) cultures. Invasion is a cellular outcome of utmost interest in cancer biology. In this protocol, we have devised an alternative strategy for evaluating cancer cell invasion in vitro, employing heterospheroids comprised of oral squamous cell carcinoma (OSCC), cancer-associated fibroblasts (CAF), and monocytes. These heterospheroids aim to mimic the tumor microenvironment (TME), including two relevant non-neoplastic cell types alongside the cancer cells. Each cell type was labeled with vital fluorescent markers emitting in distinct wavelengths before spheroid formation. Once formed, heterospheroids were seeded onto a layer of human leiomyoma-derived extracellular matrix in the upper compartment of a microporous membrane. Invasion was assessed in the z-axis using confocal microscopy. Digital images were obtained in the corresponding fluorescent channels at 10 &#181;m intervals, covering a depth of 90 &#181;m in the z-axis. Analysis was performed using freeware image software by calculating the integrated fluorescence intensity in each image and fluorescence channel. This approach enables a more dynamic analysis of cell invasion patterns in a multilayered context, as well as the examination of spatial co-localization of different cell types during invasion.

作者聚焦:破译头颈癌进展过程中癌症和免疫细胞之间的串扰

Invasion is a major biological phenomenon in the development and progression of cancer. This process is influenced by non-neoplastic cells and components of the tumor microenvironment. The purpose of this study is to describe an alternative method for analyzing tumor cell invasion in vitro using three-dimensional (3D) cultures.

分析 3D 培养物体外细胞侵袭的替代策略

Spheroid culture is a 3D model that provides an improved replication of the in vivo microenvironment compared to traditional two-dimensional (2D) ...

Three-Dimensional (3D) Tumor Spheroid Invasion Assay

Invasion of surrounding normal tissues is generally considered to be a key hallmark of malignant (as opposed to benign) tumors. For some cancers in particular (e.g., brain tumors such as glioblastoma multiforme and squamous cell carcinoma of the head and neck &#8211; SCCHN) it is a cause of severe morbidity and can be life-threatening even in the absence of distant metastases. In addition, cancers which have relapsed following treatment unfortunately often present with a more aggressive phenotype. Therefore, there is an opportunity to target the process of invasion to provide novel therapies that could be complementary to standard anti-proliferative agents. Until now, this strategy has been hampered by the lack of robust, reproducible assays suitable for a detailed analysis of invasion and for drug screening. Here we provide a simple micro-plate method (based on uniform, self-assembling 3D tumor spheroids) which has great potential for such studies. We exemplify the assay platform using a human glioblastoma cell line and also an SCCHN model where the development of resistance against targeted epidermal growth factor receptor (EGFR) inhibitors is associated with enhanced matrix-invasive potential. We also provide two alternative methods of semi-automated quantification: one using an imaging cytometer and a second which simply requires standard microscopy and image capture with digital image analysis.

Three-Dimensional 3D Tumor Spheroid Invasion Assay

Invasion of surrounding normal tissues is a defining characteristic of malignant tumors. We provide here a simple, semi-automated micro-plate assay of invasion into a natural 3D biomatrix that has been exemplified with a number of models of advanced human cancers.

三维（3D）肿瘤球体侵袭试验

Invasion of surrounding normal tissues is generally considered to be a key hallmark of malignant (as opposed to benign) tumors. For some cancers in ...

<meta charset="utf8"></head><body>检查 3D 胶原基质设置中的巨噬细胞-肿瘤球状体相互作用

A Macrophage-Tumor Spheroid Co-Invasion Assay

The interaction of immune and cancer cells and their respective impact on metastasis represents an important aspect of cancer research. So far, only a few protocols are available that allow an in vitro approximation of the in vivo situation. Here, we present a novel approach to observing the impact of human macrophages on the invasiveness of cancer cells, using tumor spheroids of H1299 non-small cell lung carcinoma cells embedded in a three-dimensional (3D) collagen I matrix. With this co-cultivation setup, we tested the impact of small interfering RNA (siRNA)-based depletion of regulatory factors in macrophages on the 3D invasion of cancer cells from the tumor spheroid compared to controls. This method allows the determination of different parameters, such as spheroid area or the number of invading cancer cells, and thus, to detect differences in cancer cell invasion. In this article, we present the respective setup, discuss the subsequent analysis, as well as the advantages and potential pitfalls of this method.

<meta charset="utf8"></head><body>巨噬细胞-肿瘤球状体共侵袭测定

Here, we present a protocol to investigate the interaction of primary human monocyte-derived macrophages with a tumor spheroid in a three-dimensional (3D) collagen I matrix, with the possibility to compare the impact of soluble and physical properties of the microenvironment on cell invasion.

巨噬细胞-肿瘤球状体共侵袭测定

The interaction of immune and cancer cells and their respective impact on metastasis represents an important aspect of cancer research. So far, only a few ...

癌细胞球体检测，以评估入侵的3D设置

摘要

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此视频中的章节

球体的检测，以测量TGF -β诱导的入侵

三维流状入侵检测中的全人肿瘤基基质

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

监测三D培养中的癌细胞入侵和T细胞毒性

分析 3D 培养物体外细胞侵袭的替代策略

分析 3D 培养物体外细胞侵袭的替代策略

分析 3D 培养物体外细胞侵袭的替代策略

三维（3D）肿瘤球体侵袭试验

巨噬细胞-肿瘤球状体共侵袭测定

巨噬细胞-肿瘤球状体共侵袭测定