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Initial contact guidance during cell spreading is contractility-independent

Abstract

A wide variety of cell types exhibit substrate topography-based behavior, also known as contact guidance. However, the precise cellular mechanisms underlying this process are still unknown. In this study, we investigated contact guidance by studying the reaction of human endothelial cells (ECs) to well-defined microgroove topographies, both during and after initial cell spreading. As the cytoskeleton plays a major role in cellular adaptation to topographical features, two methods were used to perturb cytoskeletal structures. Inhibition of actomyosin contractility with the chemical inhibitor blebbistatatin demonstrated that initial contact guidance events are independent of traction force generation. However, cell alignment to the grooved substrate was altered at later time points, suggesting an initial ‘passive’ phase of contact guidance, followed by a contractility-dependent ‘active’ phase that relies on mechanosensitive feedback. The actin cytoskeleton was also perturbed in an indirect manner by culturing cells upside down, resulting in decreased levels of contact guidance and providing evidence that a combination of gravity force and hydrostatic pressure are essential for efficient entrainment of cytoskeletal structures in topographic features. The process of contact guidance at the microscale was found to be primarily lamellipodia driven, as no bias in filopodia extension was observed on micron-scale grooves.

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Publication details

The article was received on 30 Nov 2016, accepted on 16 Jun 2017 and first published on 16 Jun 2017


Article type: Paper
DOI: 10.1039/C6SM02685K
Citation: Soft Matter, 2017, Accepted Manuscript
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    Initial contact guidance during cell spreading is contractility-independent

    A. Sales Ramos, A. Holle and R. Kemkemer, Soft Matter, 2017, Accepted Manuscript , DOI: 10.1039/C6SM02685K

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