收缩环境中细胞行为的研究与我们对细胞侵袭机制的理解特别相关。在这方面,微流体系统提供了有前途的平台,因为微制造的流体芯片提供了良好控制的物理,化学和密闭环境来研究细胞表型和行为。这里,我们报告了一种快速有效的用户友好的微流控芯片的制造过程,非常适合与免疫荧光显微镜结合进行定量活细胞分析。芯片体,聚二甲基硅氧烷,由两个培育室组成,所述培育室通过一个矩形中间进入通道连接,所述矩形中间进入通道提供到可以进行观察的一系列横向狭缝的入口。狭缝的高度被设计成略小于所研究的细胞的高度。要验证芯片性能,我们分析了在不同程度的空间限制下内皮细胞细胞骨架的重组。我们说明了狭窄的环境如何影响内皮细胞行为,以诱导podosome的形成。此外,当狭缝表面涂有纤连蛋白薄层时,该过程进一步受到刺激。该研究证明了该技术过程对于成本有效地制造用于生物应用的定制一次性芯片的适用性。
The
study of cell behavior in constricted environment is particularly relevant to our understanding of the mechanisms of cell invasion. In this regard, microfluidic systems offer promising platforms as microfabricated fluidic chips provide well-controlled physical, chemical and confined environments to
study cell phenotype and behavior. Here, we report a fast and effective manufacturing process of user-friendly microfluidic chips ideally suited for quantitative live cell analysis in combination with immunofluorescence microscopy. The chip body, made of polydimethylsiloxane, is composed of two incubation chambers connected by one rectangular intermediate entry channel which provides access to a series of transversal slits where the observation can be made. The height of the slit is designed to be slightly smaller than that of the cells under
study. To validate the chip performance, we analyzed the reorganization of the cytoskeleton of endothelial cells under various degree of spatial confinement. We illustrate how the constricted environment affects endothelial cell behavior in inducing the formation of
podosomes. Moreover, the process was stimulated further when the surface of the slit was coated with a thin layer of fibronectin. The
study demonstrates the suitability of this technological process for cost-effective fabrication of custom-made single-use chips for biological applications.