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Methods. 2016 Feb 1;94:65-74. doi: 10.1016/j.ymeth.2015.09.001. Epub 2015 Sep 2.

Microfluidic devices for the study of actin cytoskeleton in constricted environments: Evidence for podosome formation in endothelial cells exposed to a confined slit.

Author information

1
Université de Bordeaux, Bordeaux F-33000, France; INSERM U1045, Bordeaux F-33000, France.
2
Nano Science and Technology Program, Taiwan International Graduate Program, Academia Sinica, Taipei 11529, Taiwan; Institute of Physics, Academia Sinica, Taipei 11529, Taiwan; Department of Engineering and System Science, National Tsing Hua University, Hsinchu 30013, Taiwan.
3
Université de Bordeaux, Bordeaux F-33000, France.
4
Nano Science and Technology Program, Taiwan International Graduate Program, Academia Sinica, Taipei 11529, Taiwan; Institute of Physics, Academia Sinica, Taipei 11529, Taiwan; Genomics Research Center and Research Center for Applied Sciences, Academia Sinica, Taipei 11529, Taiwan. Electronic address: cfchou@phys.sinica.edu.tw.
5
Université de Bordeaux, Bordeaux F-33000, France; INSERM U1045, Bordeaux F-33000, France. Electronic address: elisabeth.genot@inserm.fr.

Abstract

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.

KEYWORDS:

Actin cytoskeleton; Confined environment; Invasion; Microfluidics; Migration; Podosomes

PMID:
26342258
DOI:
10.1016/j.ymeth.2015.09.001
[Indexed for MEDLINE]

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