Microfluidic Collective Cell Migration Assay for Study of Endothelial Cell Proliferation and Migration under Combinations of Oxygen Gradients, Tensions, and Drug Treatments
Microfluidic Collective Cell Migration Assay for Study of Endothelial Cell Proliferation and Migration under Combinations of Oxygen Gradients, Tensions, and Drug Treatments
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DOI：
10.1038/s41598-019-44594-5
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发表时间：
2019-06-03
影响因子：
4.6
通讯作者：
Tung, Yi-Chung
中科院分区：
文献类型：
--
作者：
Shih, Hsiu-Chen;Lee, Tse-Ang;Tung, Yi-Chung
内皮细胞的增殖和迁移在许多生物学活动中起着重要作用，它们受到多种微环境因素的调控。在本文中，一种新的微流控集体细胞迁移实验的发展，研究内皮细胞迁移和增殖的组合下的三种氧条件：常氧，氧梯度，缺氧和三种培养基成分：正常生长培养基，培养基与细胞松弛素-D的肌动蛋白聚合抑制，和YC-1的缺氧诱导因子（HIF）抑制。本文设计的微流控装置允许使用层流图案化形成具有一致尺寸的细胞图案。此外，可以通过空间受限的化学反应方法在装置内产生稳定的氧梯度。该装置可以在传统的细胞培养箱中操作，具有最少的化学试剂和仪器，用于实际应用。结果显示，对于所有培养基组合物，在氧梯度下内皮细胞的定向集体细胞迁移。定向行为以前从未讨论过，并且表明氧梯度在各种生物活动期间引导内皮细胞迁移中的关键作用。该方法为进一步研究内皮细胞在各种生理微环境下的行为提供了一个实用而有力的工具。
Proliferation and migration of endothelial cells play an important role in many biological activities, and they can be regulated by various microenvironmental factors. In this paper, a novel microfluidic collective cell migration assay is developed to study endothelial cell migration and proliferation under combinations of three oxygen conditions: normoxia, oxygen gradient, and hypoxia and three medium compositions: normal growth medium, the medium with cytochalasin-D for actin polymerization inhibition, and with YC-1 for hypoxia-inducible factor (HIF) inhibition. The microfluidic device designed in the paper allows cell patterns formed with consistent dimensions using laminar flow patterning. In addition, stable oxygen gradients can be generated within the device by a spatially confined chemical reaction method. The device can be operated in conventional cell incubators with minimal chemical reagents and instrumentation for practical applications. The results show directional collective cell migration of the endothelial cells under the oxygen gradients for all the medium compositions. The directional behavior has never been discussed before, and indicates critical roles of oxygen gradients in guiding endothelial cell migration during various biological activities. The developed assay provides a practical yet powerful tool for further in vitro study of endothelial cell behaviors under various physiological microenvironments.