Abstract
BackgroundIn this work, we explore how U2OS cells are affected by arrays of polymer nanopillars fabricated on flat glass surfaces. We focus on describing changes to the organisation of the actin cytoskeleton and in the location, number and shape of focal adhesions. From our findings we identify that the cells can be categorised into different regimes based on their spreading and adhesion behaviour on nanopillars. A quantitative analysis suggests that cells seeded on dense nanopillar arrays are suspended on top of the pillars with focal adhesions forming closer to the cell periphery compared to flat surfaces or sparse pillar arrays. This change is analogous to similar responses for cells seeded on soft substrates.ResultsIn this work, we explore how U2OS cells are affected by arrays of polymer nanopillars fabricated on flat glass surfaces. We focus on describing changes to the organisation of the actin cytoskeleton and in the location, number and shape of focal adhesions. From our findings we identify that the cells can be categorised into different regimes based on their spreading and adhesion behaviour on nanopillars. A quantitative analysis suggests that cells seeded on dense nanopillar arrays are suspended on top of the pillars with focal adhesions forming closer to the cell periphery compared to flat surfaces or sparse pillar arrays. This change is analogous to similar responses for cells seeded on soft substrates.ConclusionOverall, we show that the combination of high throughput nanofabrication, advanced optical microscopy, molecular biology tools to visualise cellular processes and data analysis can be used to investigate how cells interact with nanostructured surfaces and will in the future help to create culture substrates that induce particular cell function.Graphic
Highlights
In this work, we explore how U2OS cells are affected by arrays of polymer nanopillars fabricated on flat glass surfaces
One approach is to use flat-surfaces decorated with vertically aligned nanostructures as a simple model system
High resolution live cell imaging of co-transfected U2OS cells expressing pCMV-LifeActGFP and pTAGRFP-Vinculin have been used to study the influence of nanopillar arrays on actin cytoskeleton focal adhesion organisation
Summary
We explore how U2OS cells are affected by arrays of polymer nanopillars fabricated on flat glass surfaces. A quantitative analysis suggests that cells seeded on dense nanopillar arrays are suspended on top of the pillars with focal adhesions forming closer to the cell periphery compared to flat surfaces or sparse pillar arrays This change is analogous to similar responses for cells seeded on soft substrates. The ECM serves as a structural scaffold for the cells, it is a conveyor of biomechanical and biochemical signals and regulates a range of processes such as tissue morphogenesis, homeostatis and differentiation It is Motivated by the need of creating cell culturing models that better represent in vivo conditions, researchers have increasingly started to study cell behaviour in 3D matrices and in “semi-3D” systems. Characteristics such as viability, proliferation, differentiation and morphology are known to differ between cells on flat surfaces and cells embedded in 3D matricies [3, 7].
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