Evaluation of cancer stem cell migration using compartmentalizing microfluidic devices and live cell imaging.
Evaluation of cancer stem cell migration using compartmentalizing microfluidic devices and live cell imaging.
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DOI：
10.3791/3297
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发表时间：
2011-12-23
期刊：
影响因子：
--
通讯作者：
Kuo JS
中科院分区：
文献类型：
--
作者：
Huang Y;Agrawal B;Clark PA;Williams JC;Kuo JS
过去40年，美国在癌症研究上投入了超过2000亿美元，结果死亡率仅下降了5%。改善患者治疗效果的一个主要障碍是对与侵袭性癌细胞侵袭、转移和治疗耐药相关的细胞迁移机制了解甚少。多形性胶质母细胞瘤（GBM）是最常见的原发性恶性成人脑肿瘤2，就体现了这种困难。尽管进行了标准手术、放疗和化疗，但由于 GBM 侵袭邻近大脑且癌症快速复发，患者的中位生存期仅为 15 个月2。异常细胞迁移机制和肿瘤微环境的相互作用可能将癌症与正常细胞区分开来。因此，改善 GBM 的治疗方法需要更好地了解癌细胞迁移机制。最近的研究表明，GBM 内的一小部分细胞，即脑肿瘤干细胞 (BTSC)，可能是治疗耐药和复发的原因。 BTSC 迁移能力的潜在机制才刚刚开始被表征1,4。由于目视检查和几何操作的限制，传统的迁移测定5仅限于量化总体细胞群。相比之下，微流体装置由于与现代显微镜兼容并可控制微环境，因此允许进行单细胞分析6-9。我们提出了一种使用分隔微流体装置详细表征 BTSC 迁移的方法。这些 PDMS 制造的设备将组织培养环境分为三个相连的隔室：接种室、接收室和桥接微通道。我们定制了该设备，使两个腔室都能容纳足够的介质，以支持可行的 BTSC 4-5 天，而无需更换介质。最初引入接种室的高度移动的 BTSC 在迁移后通过桥接微通道与平行接收室隔离。这种迁移模拟癌细胞通过大脑间隙的扩散。迁移过程中细胞形态的实时相位图像被记录数天。因此，可以对高度迁移的 BTSC 进行分离、重新培养和进一步分析。 区室化微流体可以成为研究 BTSC 和其他癌症干细胞迁移行为的多功能平台。通过结合梯度发生器、流体处理、微电极和其他微流体模块，这些设备还可用于药物筛选和疾病诊断6。分离迁移细胞的侵袭性亚群将使研究潜在的分子机制成为可能。
In the last 40 years, the United States invested over 200 billion dollars on cancer research, resulting in only a 5% decrease in death rate. A major obstacle for improving patient outcomes is the poor understanding of mechanisms underlying cellular migration associated with aggressive cancer cell invasion, metastasis and therapeutic resistance1. Glioblastoma Multiforme (GBM), the most prevalent primary malignant adult brain tumor2, exemplifies this difficulty. Despite standard surgery, radiation and chemotherapies, patient median survival is only fifteen months, due to aggressive GBM infiltration into adjacent brain and rapid cancer recurrence2. The interactions of aberrant cell migratory mechanisms and the tumor microenvironment likely differentiate cancer from normal cells3. Therefore, improving therapeutic approaches for GBM require a better understanding of cancer cell migration mechanisms. Recent work suggests that a small subpopulation of cells within GBM, the brain tumor stem cell (BTSC), may be responsible for therapeutic resistance and recurrence. Mechanisms underlying BTSC migratory capacity are only starting to be characterized1,4. Due to a limitation in visual inspection and geometrical manipulation, conventional migration assays5 are restricted to quantifying overall cell populations. In contrast, microfluidic devices permit single cell analysis because of compatibility with modern microscopy and control over micro-environment6-9. We present a method for detailed characterization of BTSC migration using compartmentalizing microfluidic devices. These PDMS-made devices cast the tissue culture environment into three connected compartments: seeding chamber, receiving chamber and bridging microchannels. We tailored the device such that both chambers hold sufficient media to support viable BTSC for 4-5 days without media exchange. Highly mobile BTSCs initially introduced into the seeding chamber are isolated after migration though bridging microchannels to the parallel receiving chamber. This migration simulates cancer cellular spread through the interstitial spaces of the brain. The phase live images of cell morphology during migration are recorded over several days. Highly migratory BTSC can therefore be isolated, recultured, and analyzed further. Compartmentalizing microfluidics can be a versatile platform to study the migratory behavior of BTSCs and other cancer stem cells. By combining gradient generators, fluid handling, micro-electrodes and other microfluidic modules, these devices can also be used for drug screening and disease diagnosis6. Isolation of an aggressive subpopulation of migratory cells will enable studies of underlying molecular mechanisms.
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影响因子：
158.5
作者：
Stupp, R;Mason, WP;Ryan, G
通讯作者：
Ryan, G
影响因子：
6.1
作者：
Millet, Larry J.;Stewart, Matthew E.;Gillette, Martha U.
通讯作者：
Gillette, Martha U.
影响因子：
3
作者：
Svendsen, CN;ter Borg, MG;Caldwell, MA
通讯作者：
Caldwell, MA
影响因子：
50.3
作者：
Chen, Ruihuan;Nishimura, Merry C.;Phillips, Heidi S.
通讯作者：
Phillips, Heidi S.
影响因子：
11.2
作者：
Galli, R;Binda, E;Vescovi, A
通讯作者：
Vescovi, A