On-chip human microvasculature assay for visualization and quantification of tumor cell extravasation dynamics.
On-chip human microvasculature assay for visualization and quantification of tumor cell extravasation dynamics.
复制标题
DOI：
10.1038/nprot.2017.018
复制
发表时间：
2017-05
期刊：
影响因子：
14.8
通讯作者：
Kamm RD
中科院分区：
文献类型：
--
作者：
Chen MB;Whisler JA;Fröse J;Yu C;Shin Y;Kamm RD
导致>90%的癌症相关死亡的远处转移是通过肿瘤细胞经由循环的血行性传播实现的。这需要完成一系列复杂的步骤，包括过境、初始滞留、外渗、存活和扩散。增加对这些步骤中每一个步骤的细胞和分子参与者的理解是发现早期转移性播散期间治疗干预的新机会的关键。在这里，我们描述了一个体外模型的人微循环与潜在的重演离散步骤的早期转移播种，包括逮捕，跨内皮迁移和早期微转移形成。该微型装置具有在4-5天内形成的自组织人类微血管网络，之后可以通过标准共聚焦显微镜灌注肿瘤并在72小时内轻松跟踪外渗事件。与大多数体内和体外外渗测定相反，由于将血管网络限制在靠近装置表面的一个平面上，因此可以容易地实现血管外细胞的稳健和快速评分与高分辨率成像的组合。这使得血管外细胞明显不同，并允许与厚组织中的肿瘤细胞相比快速识别感兴趣的肿瘤细胞。每个实验生成大量器械（约36个）的能力与快速定量相结合，进一步允许进行高度参数化研究，这在测试多种遗传或药理学扰动时是必需的。这与实时跟踪单细胞外渗事件的能力相结合，允许用中等数量的数据点高度详细地观察肿瘤和内皮形态学动态。该协议扩展描述了现有协议的改编，该协议描述了提供额外应用的微流体平台。
Distant metastasis, which results in >90% of cancer related deaths, is enabled by hematogenous dissemination of tumor cells via the circulation. This requires the completion of a sequence of complex steps including transit, initial arrest, extravasation, survival and proliferation. Increased understanding of the cellular and molecular players enabling each of these steps is key in uncovering new opportunities for therapeutic intervention during early metastatic dissemination. Here, we describe an in vitro model of the human microcirculation with the potential to recapitulate discrete steps of early metastatic seeding, including arrest, transendothelial migration and early micrometastases formation. The microdevice features self-organized human microvascular networks formed over 4–5 days, after which tumor can be perfused and extravasation events easily tracked over 72 hours, via standard confocal microscopy. Contrary to most in vivo and in vitro extravasation assays, robust and rapid scoring of extravascular cells combined with high-resolution imaging can be easily achieved due to the confinement of the vascular network to one plane close to the surface of the device. This renders extravascular cells clearly distinct and allows tumor cells of interest to be identified quickly compared to those in thick tissues. The ability to generate large numbers of devices (~36) per experiment coupled with fast quantitation further allows for highly parametric studies, which is required when testing multiple genetic or pharmacological perturbations. This is coupled with the capability for live tracking of single cell extravasation events allowing both tumor and endothelial morphological dynamics to be observed in high detail with a moderate number of data points. This Protocol Extension describes an adaptation of an existing Protocol describing a microfluidic platform that offers additional applications.
登录
查看更多内容
DOI：
10.1039/c3ib40170g
发表时间：
2014-06
期刊：
Integrative biology : quantitative biosciences from nano to macro
影响因子：
--
作者：
Ehsan SM;Welch-Reardon KM;Waterman ML;Hughes CC;George SC
通讯作者：
George SC
影响因子：
82.9
作者：
通讯作者：
--
影响因子：
3.2
作者：
Kim, Jin;Yang, Kisuk;Lee, Jun Hyup
通讯作者：
Lee, Jun Hyup
影响因子：
21.3
作者：
通讯作者：
--
影响因子：
50.3
作者：
Labelle M;Begum S;Hynes RO
通讯作者：
Hynes RO