Adsorption state of fibronectin on poly(dimethylsiloxane) surfaces with varied stiffness can dominate adhesion density of fibroblasts

Seo, Ji Hun; Sakai, Keiko; Yui, Nobuhiko

doi:10.1016/j.actbio.2012.10.015

Cited by 68 publications

(69 citation statements)

References 30 publications

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“…Recent evidence suggests that cells react to a complex mechanosensing apparatus whereby the interaction of ligand tethering and ECM stiffness can impart differential cellular functions [18], including differentiation [19]. Indeed, emerging reports have highlighted the significance of protein conformational status which is influenced by the underlying matrix stiffness [20], or chemistry [21] effectively modulating the presentation of binding sites for cell receptors and/or growth factors [22], the entirety of which can significantly influence cellular processes [23].…”

Section: Discussionmentioning

confidence: 99%

Three-dimensional, soft neotissue arrays as high throughput platforms for the interrogation of engineered tissue environments

Floren

Tan

2015

Biomaterials

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Local signals from tissue-specific extracellular matrix (ECM) microenvironments, including matrix adhesive ligand, mechanical elasticity and micro-scale geometry, are known to instruct a variety of stem cell differentiation processes. Likewise, these signals converge to provide multifaceted, mechanochemical cues for highly-specific tissue morphogenesis or regeneration. Despite accumulated knowledge about the individual and combined roles of various mechanochemical ECM signals in stem cell activities on 2-dimensional matrices, the understandings of morphogenetic or regenerative 3-dimenstional tissue microenvironments remain very limited. To that end, we established high-throughput platforms based on soft, fibrous matrices with various combinatorial ECM proteins meanwhile highly-tunable in elasticity and 3-dimensional geometry. To demonstrate the utility of our platform, we evaluated 64 unique combinations of 6 ECM proteins (collagen I, collagen III, collagen IV, laminin, fibronectin, and elastin) on the adhesion, spreading and fate commitment of mesenchymal stem cell (MSCs) under two substrate stiffness (4.6 kPa, 20 kPa). Using this technique, we identified several neotissue microenvironments supporting MSC adhesion, spreading and differentiation toward early vascular lineages. Manipulation of the matrix properties, such as elasticity and geometry, in concert with ECM proteins will permit the investigation of multiple and distinct MSC environments. This paper demonstrates the practical application of high through-put technology to facilitate the screening of a variety of engineered microenvironments with the aim to instruct stem cell differentiation.

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Section: Discussionmentioning

confidence: 99%

Three-dimensional, soft neotissue arrays as high throughput platforms for the interrogation of engineered tissue environments

Floren

Tan

2015

Biomaterials

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“…One of the primary advantages of PDMS over substrates such as glass and polystyrene is its tunable mechanical nature and flexibility. However, to date, most studies involving changes in stiffness of PDMS used un-patterned surfaces [11, 28, 30]. Here, in order to optimize cell micropatterning for different cell types, underlying substrate stiffness was further tested as an additional cue.…”

Section: Resultsmentioning

confidence: 99%

“…Despite being less stiff at the macroscale, 20:1 PDMS is more pliable at the nanoscale resulting in exposed cell-binding motifs [30]. On the other hand, a different study found transformed fibroblasts to grow at similar rates on PDMS irrespective of stiffness [11].…”

Section: Resultsmentioning

confidence: 99%

“…For instance, our group and others have previously shown that human embryonic stem cell proliferation can be affected by varying the stiffness [28, 29]. Characteristically, exposure of cell-binding motifs differ based on the nanoscale surface stiffness, reflecting changes in cell behavior [30]. In this work, we investigate the photopatterning of PEG-DA hydrogels that can be used on PDMS surfaces with different substrate stiffness.…”

Section: Introductionmentioning

confidence: 99%

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Large area micropatterning of cells on polydimethylsiloxane surfaces

et al. 2014

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BackgroundPrecise spatial control and patterning of cells is an important area of research with numerous applications in tissue engineering, as well as advancing an understanding of fundamental cellular processes. Poly (dimethyl siloxane) (PDMS) has long been used as a flexible, biocompatible substrate for cell culture with tunable mechanical characteristics. However, fabrication of suitable physico-chemical barriers for cells on PDMS substrates over large areas is still a challenge.ResultsHere, we present an improved technique which integrates photolithography and cell culture on PDMS substrates wherein the barriers to cell adhesion are formed using the photo-activated graft polymerization of polyethylene glycol diacrylate (PEG-DA). PDMS substrates with varying stiffness were prepared by varying the base to crosslinker ratio from 5:1 to 20:1. All substrates show controlled cell attachment confined to fibronectin coated PDMS microchannels with a resistance to non-specific adhesion provided by the covalently immobilized, hydrophilic PEG-DA.ConclusionsUsing photolithography, it is possible to form patterns of high resolution stable at 37°C over 2 weeks, and microstructural complexity over large areas of a few cm2. As a robust and scalable patterning method, this technique showing homogenous and stable cell adhesion and growth over macroscales can bring microfabrication a step closer to mass production for biomedical applications.Electronic supplementary materialThe online version of this article (doi:10.1186/1754-1611-8-24) contains supplementary material, which is available to authorized users.

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“…Factors that demonstrate significant influence on cell adhesion include substrate mechanical properties [1] [2] [3], the type of adhesive ligands [4] [5] , and the spatial characteristics of ligand presentation [6]. Control over multiple design characteristics is required in order to reach the complexity of the natural and three-dimensional ECM.…”

Section: Introductionmentioning

confidence: 99%