Micropatterning Strategies to Engineer Controlled Cell and Tissue Architecture in Vitro

D’Arcangelo, Elisa; McGuigan, Alison P.

doi:10.2144/000114245

Cited by 76 publications

(57 citation statements)

References 71 publications

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“…The aim of this investigation is to elucidate the general biophysical mechanisms underlying contact guidance of individual cells. Various in-vitro chemical micropatterning approaches have been developed to study cellular contact guidance, as model systems to simplify the highly complex in vivo environments [19,20]. We followed such an approach and seeded a low density of myofibroblasts (specifically, Human Vena Saphena Cells; see Methods) on effectively rigid substrates microprinted with fibronectin stripes of widths , ranging from greater than 1.5 mm (resembling a homogeneous 2D substrate) to 50 μm; see Fig.…”

Section: Introductionmentioning

confidence: 99%

Entropic forces drive cellular contact guidance

Buskermolen

Suresh²,

Shishvan

et al. 2018

Preprint

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Contact guidance-the widely-known phenomenon of cell alignment induced by anisotropic environmental features-is an essential step in the organization of adherent cells, but the mechanisms by which cells achieve this orientational ordering remain unclear. Here we seeded myofibroblasts on substrates micropatterned with stripes of fibronectin and observed that contact guidance emerges at stripe widths much greater than the cell size. To understand the origins of this surprising observation, we combined morphometric analysis of cells and their subcellular components with a novel statistical framework for modelling non-thermal fluctuations of living cells. This modelling framework is shown to predict not only the trends but also the statistical variability of a wide range of biological observables including cell (and nucleus) shapes, sizes and orientations, as well as stress-fibre arrangements within the cells with remarkable fidelity. By comparing observations and theory, we identified two regimes of contact guidance: (i) guidance on stripe widths smaller than the cell size ( ≤ 160 μm), which is accompanied by biochemical changes within the cells, including increasing stress-fibre polarisation and cell elongation, and (ii) entropic guidance on larger stripe widths, which is governed by fluctuations in the cell morphology. Overall, our findings suggest an entropymediated mechanism for contact guidance associated with the tendency of cells to maximise their morphological entropy through shape fluctuations.

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Section: Introductionmentioning

confidence: 99%

Entropic forces drive cellular contact guidance

Buskermolen

Suresh²,

Shishvan

et al. 2018

Preprint

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“…This can be achieved by creating a nonadhesive area via a physical barrier (using stamps, as seen in (21)) or a chemical barrier (e.g., bovine serum albumin (BSA) (20)). The first population of cells will adhere to the area surrounding the barrier; once the barrier is removed (either by physical removal or by coating the nonadhesive area with an adhesive matrix factor (20)), the second cell population binds in the remaining space (22). This method has the advantage of allowing cells to interact via both secreted factors and cellcell interactions, but is laborious, requiring additional steps and specialized equipment.…”

mentioning

confidence: 99%

A Triple Co-Culture Method to Investigate the Effect of Macrophages and Fibroblasts on Myoblast Proliferation and Migration

Venter

Niesler

2018

BioTechniques

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The communication between nonmyogenic cells, such as macrophages and fibroblasts, and myoblasts is crucial for successful skeletal muscle repair. In vitro co-culture methods can be used to increase our understanding of these cellular interactions; however, current protocols are restricted to two, often physically separate, cell populations. Here, we demonstrate a novel, inexpensive in vitro triple co-culture method that facilitates the co-culture of at least three cell populations with some degree of cell–cell contact. Using this method, we determined the effect of macrophages and fibroblasts on myoblast proliferation and migration. A significant increase in myoblast proliferation and migration was observed following co-culture with either macrophages or fibroblasts. However, triple co-culture of macrophages, fibroblasts, and myoblasts revealed that the presence of macrophages prevented fibroblasts from maintaining this positive effect on myoblast migration. Macrophages, on the other hand, continued to promote myoblast proliferation whether in the presence of fibroblasts or not. Our triple co-culture system highlights the significance of multicellular communication in regulating myoblast proliferation and migration and emphasizes the importance of more complex co-culture systems when investigating myogenesis in vitro.

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“…For example, grinding offers a smaller potential feature size, due to it not being limited by resolution, or spot size as the case with photolithography [16] and laser processing [10], respectively. It is also a relatively simpler manufacturing process, not requiring a clean room as is the case with lithography [6]. Little research has been done on grinding micro-patterned surfaces for cell control.…”

Section: Introductionmentioning

confidence: 99%

Machine grinding as an alternative method for creating functional surfaces for controlling cell behaviour

Irving

Murphy

Morgan

et al. 2016

Int J Adv Manuf Technol

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There is extensive evidence to show that certain cellular behaviours including cell proliferation, migration and adhesion can be controlled by culturing cells on surfaces containing different micro-metre-and nanometre-scale features. This paper will introduce the use of machine grinding to generate surfaces with micro-sized features and their ability to affect cell behaviour. Results are presented which show that polyurethane castings of the ground surfaces can promote cell adhesion and migration. This study demonstrates the usefulness of surface grinding as a cost-effective method for generating functional surfaces for modifying cell behaviour.

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Micropatterning Strategies to Engineer Controlled Cell and Tissue Architecture in Vitro

Cited by 76 publications

References 71 publications

Entropic forces drive cellular contact guidance

Entropic forces drive cellular contact guidance

A Triple Co-Culture Method to Investigate the Effect of Macrophages and Fibroblasts on Myoblast Proliferation and Migration

Machine grinding as an alternative method for creating functional surfaces for controlling cell behaviour

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