Research Center for Bioengineering and Sensing Technology, School of Chemistry and Biological Engineering, Institute of Precision Medicine and Health, Beijing Key Laboratory for Bioengineering and Sensing Technology, University of Science and Technology Beijing , Beijing 100083, China.
Anal Chem. 2018 Jan 2;90(1):777-784. doi: 10.1021/acs.analchem.7b03225. Epub 2017 Dec 18.
Cancer metastatic dissemination is a complex event during tumor progression which involves cell-cell and cell-matrix interactions. Micropatterning is one of the most efficient ways to study tumor development because it can tune the distribution of cells with spatial and temporal control. Extensive studies have shown that microfluidics can provide a feasible method for cell patterning. However, the current technique requires a microfabrication laboratory to manufacture the chip, which results in inaccessibility to researchers, especially biologists who focus on disclosing biological mechanisms rather than the methods. In this work, we developed a new methodology (tape-assisted photolithographic-free microfluidic chip, TAPMiC) that can realize homogeneous and heterogeneous micropatterning (45 features, 300 μm diameter of each) on a culture dish without the photolithographic procedure. We have applied this method to study critical biological problems, such as tumor cell migration under different conditions, including antitumor pharmaceutics and candidate gene RNAi assay that was relevant to tumor translocation and invasion. Moreover, this platform can achieve copatterning to recapitulate the tumor invasion scenario with single-cell trackable analysis. To decode regulation during metastasis, we conducted in situ recovering for quantitative polymerase chain reaction (qPCR) analysis from each cell type from tumor-fibroblast copairing. Regulation of several essential genes has unveiled that matrix degradation gene MMP2 and angiogenesis associated gene VEGFA were up-regulated in tumor cells in the fibroblast-enriched niche compared with homogeneous cultivation. Therefore, this approach constitutes a novel tool for investigating metastasis with quantitative measurements both on phenotype and genetical information.
肿瘤转移扩散是肿瘤进展过程中的一个复杂事件，涉及细胞-细胞和细胞-基质相互作用。微图案化是研究肿瘤发展的最有效方法之一，因为它可以通过时空控制来调整细胞的分布。大量研究表明，微流控技术可以为细胞图案化提供一种可行的方法。然而，目前的技术需要一个微制造实验室来制造芯片，这使得研究人员，特别是专注于揭示生物学机制而不是方法的生物学家无法使用。在这项工作中，我们开发了一种新的方法（胶带辅助免光刻微流控芯片，TAPMiC），可以在培养皿上实现均匀和异质的微图案化（45 个特征，每个特征直径为 300μm），而无需光刻程序。我们已经应用这种方法来研究关键的生物学问题，例如肿瘤细胞在不同条件下的迁移，包括抗肿瘤药物和候选基因 RNAi 测定，这与肿瘤转移和侵袭有关。此外，该平台可以实现共图案化，以单细胞可追踪分析再现肿瘤侵袭场景。为了解码转移过程中的调控，我们对来自肿瘤-成纤维细胞共培养的每个细胞类型进行了原位恢复的定量聚合酶链反应（qPCR）分析。对几个重要基因的调控发现，与同质培养相比，基质降解基因 MMP2 和血管生成相关基因 VEGFA 在富含成纤维细胞的生态位中的肿瘤细胞中上调。因此，这种方法构成了一种新的工具，可以对表型和遗传信息进行定量测量，从而研究转移。
