| Single Cell Migration Assay Using Human Breast Cancer MDA-MB-231 Cell Line
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Author:
Date:
2020-04-20
[Abstract] Cell migration is a fundamental cellular process that plays a crucial role in many physioglogical and pathological processes such as wound healing or cancer metastasis. Many assays have been developed to examine cell migration, such as the wound healing or scratch assay, Boyden Chamber or transwell assay, and the method we will describe here, single cell migration assay. In this assay, cells are plated sparsely on a collagen coated plate and live cell imaging is performed over a period of 2 h at 1 frame per minute. After imaging is completed, cells are tracked manually using ImageJ by tracking movement of the centroid of the cell. These data points are then exported and overall distance travelled from frame to frame is determined and divided by total time imaged to determine speed of the ...
[摘要] [摘要 ] 细胞迁移是一个基本的细胞过程，在许多生理和病理过程中（如伤口愈合或癌症转移）起着至关重要的作用，已经开发了许多检测细胞迁移的方法，例如伤口愈合或刮擦实验，Boyden Chamber或Transwell 分析，我们将在此介绍的方法是单细胞迁移分析。在此分析中，将细胞稀疏地铺在胶原蛋白包被的板上，并在2小时内进行活细胞成像 成像完成后，通过跟踪细胞质心的运动使用ImageJ手动跟踪细胞，然后导出这些数据点，确定帧与帧之间的总距离并除以成像的总时间确定细胞的速度。此方法提供了一种在进行更复杂的测定之前检查细胞操作对细胞迁移影响的快速方法。
[背景 ] 细胞迁移在从胚胎发育到血管生成和肿瘤转移的生理和病理过程中均起着重要作用（Le Clainche 和Carlier ，2008）。细胞运动是高度精心策划的事件，可以概括为四个基本步骤的循环：我）膜伸出驱动下肌动蛋白聚合，二）稳定突出通过整合素介导的细胞-基质粘附，III）细胞体易位驱动通过肌动球蛋白收缩力，最后，IV）后发布结果的Mechan 的iCal力和/或细胞基质粘附组分的蛋白水解作用（Sheetz 等，1999 ; Ridley 等，2003; Panetti 等，2004; Stradal and Scita ，2006; Tomasevic 等，2007; Le Clainche and ...
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| In vitro Flow Adhesion Assay for Analyzing Shear-resistant Adhesion of Metastatic Cancer Cells to Endothelial Cells
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Author:
Date:
2016-02-20
[Abstract] Hematogenous metastasis is a primary cause of mortality from metastatic cancer. The shear-resistant adhesion of circulating tumor cells to the vascular endothelial cell surface under blood flow is an essential step in cell extravasation and further tissue invasion. This is similar to a process exploited by leukocytes for adhesion to inflamed blood vessels (leukocyte mimicry). The shear resistant adhesion is mediated by high affinity interactions between endothelial adhesion molecules and their counter receptor ligand expressed on circulating cells. Thus, weak interaction results in a rapid detachment of circulating cells from endothelium. Despite the critical role of vascular adhesion of cancer cells in hematogenous metastasis, our knowledge regarding this process has been limited due to ...
[摘要] 血液转移是转移性癌症死亡的主要原因。循环肿瘤细胞对血流内的血管内皮细胞表面的剪切粘附是细胞外渗和进一步组织侵入的必要步骤。这类似于白细胞用于粘附到发炎血管的方法（白细胞模拟）。剪切抗性粘附由内皮粘附分子和其在循环细胞上表达的其受体配体之间的高亲和力相互作用介导。因此，弱相互作用导致循环细胞从内皮快速分离。尽管癌细胞在血源性转移中的血管粘附的关键作用，我们关于该过程的知识由于难以在体外模拟动态流动条件而受到限制。为了更好地了解癌细胞对内皮的剪切抗性粘附，我们开发了一种用于测量在生理流动条件下循环肿瘤细胞对内皮细胞的剪切抗性粘附的方案，通过适应良好确立的流动粘附测定炎症细胞。该技术可用于评价1）癌细胞的抗剪切粘附能力和2）支持癌细胞粘附所必需的内皮粘附分子（Kang等人，2015）。
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| Tumor Cell Invasion Assay
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Author:
Date:
2012-02-05
[Abstract] Cell invasion assays have been used to study the interactions between tumor cells and the extracellular matrix (ECM), which not only provides a structural scaffold for the cell, but also contains various biological factors for the survival and growth of the cell. ECM gel contains the basic components of the ECM that provides a structural support for the cell to grow and move. Cells can secrete enzymes that degrade certain components of the ECM to move towards chemoattractants, or to simply establish niches for growth. Metastatic tumor cells often show more invasiveness to the ECM gel due to their higher motility and/or enzymatic activity for degrading ECM components. This protocol describes a tumor cell invasion assay to study the interactions between tumor cells and the ECM.
[摘要] 细胞侵袭测定已经用于研究肿瘤细胞和细胞外基质（ECM）之间的相互作用，其不仅为细胞提供结构支架，而且还包含用于细胞存活和生长的各种生物因子。 ECM凝胶含有ECM的基本组分，为细胞生长和移动提供结构支持。 细胞可分泌降解ECM的某些组分以趋向化学引诱物的酶，或简单地建立用于生长的壁龛。 转移性肿瘤细胞通常对ECM凝胶显示更多的侵袭性，这是由于它们对于降解ECM组分的较高的运动性和/或酶活性。 该协议描述了肿瘤细胞侵袭实验，以研究肿瘤细胞和ECM之间的相互作用。
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