Characterization of three-dimensional cancer cell migration in mixed collagen-Matrigel scaffolds using microfluidics and image analysis.
Characterization of three-dimensional cancer cell migration in mixed collagen-Matrigel scaffolds using microfluidics and image analysis.
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DOI：
10.1371/journal.pone.0171417
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Ortiz-de-Solorzano C
中科院分区：
文献类型：
--
作者：
Anguiano M;Castilla C;Maška M;Ederra C;Peláez R;Morales X;Muñoz-Arrieta G;Mujika M;Kozubek M;Muñoz-Barrutia A;Rouzaut A;Arana S;Garcia-Aznar JM;Ortiz-de-Solorzano C
微流控设备正在成为在体外重现细胞和组织行为的主流工具。在这项研究中，我们使用填充了混合胶原-Matrigel水凝胶的微流控装置来研究肺癌细胞在不同肿瘤侵袭微环境下的迁移。本文介绍了微流控装置的设计，对水凝胶的形态和力学特性进行了表征，并用定量图像分析方法测量了H1299肺腺癌细胞在不同实验条件下的迁移。我们的结果显示了肺癌细胞迁移的可塑性，从单纯的胶原基质中的间质转移到胶原-基质基质中的叶状基质，这种基质近似于破坏的基底膜和下面的结缔组织之间的界面。我们对迁移速度的量化证实了Matrigel的两相作用。在低浓度时，Matrigel促进迁移，很可能是通过提供支持和生长因子保留环境。在高浓度时，Matrigel会减缓迁移速度，这可能是由于过度附着所致。最后，我们证明了基于抗体的整合素阻断促进了从间充质或叶向阿米巴的迁移表型的变化，并分析了这种变化对迁移动力学的影响，涉及到基质的结构。总之，我们描述并表征了一个健壮的微流控平台和一套可用于研究不同微环境和实验条件下肺癌细胞迁移的软件工具。该平台可用于未来的研究，从而受益于微流控设备带来的优势：环境的精确控制、优异的光学特性、高通量研究的并行化以及治疗药物的高效使用。
Microfluidic devices are becoming mainstream tools to recapitulate in vitro the behavior of cells and tissues. In this study, we use microfluidic devices filled with hydrogels of mixed collagen-Matrigel composition to study the migration of lung cancer cells under different cancer invasion microenvironments. We present the design of the microfluidic device, characterize the hydrogels morphologically and mechanically and use quantitative image analysis to measure the migration of H1299 lung adenocarcinoma cancer cells in different experimental conditions. Our results show the plasticity of lung cancer cell migration, which turns from mesenchymal in collagen only matrices, to lobopodial in collagen-Matrigel matrices that approximate the interface between a disrupted basement membrane and the underlying connective tissue. Our quantification of migration speed confirms a biphasic role of Matrigel. At low concentration, Matrigel facilitates migration, most probably by providing a supportive and growth factor retaining environment. At high concentration, Matrigel slows down migration, possibly due excessive attachment. Finally, we show that antibody-based integrin blockade promotes a change in migration phenotype from mesenchymal or lobopodial to amoeboid and analyze the effect of this change in migration dynamics, in regards to the structure of the matrix. In summary, we describe and characterize a robust microfluidic platform and a set of software tools that can be used to study lung cancer cell migration under different microenvironments and experimental conditions. This platform could be used in future studies, thus benefitting from the advantages introduced by microfluidic devices: precise control of the environment, excellent optical properties, parallelization for high throughput studies and efficient use of therapeutic drugs.