Use of micropatterned adhesive surfaces for control of cell behavior

LeDuc, Philip; Ostuni, Emanuele; Whitesides, George M.; Ingber, Donald E.

doi:10.1016/s0091-679x(02)69024-7

Cited by 33 publications

(25 citation statements)

References 33 publications

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“…The technological details and biological applications of µCP have been extensively discussed elsewhere (LeDuc et al, 2002;Mrksich and Whitesides, 1996). In summary, µCP is a versatile method for producing geometrically defined patterns of a variety of proteins.…”

Section: Discussionmentioning

confidence: 99%

Cell behaviour on micropatterned substrata: limits of extracellular matrix geometry for spreading and adhesion

Lehnert

Wehrle-Haller²,

Dávid

et al. 2004

Journal of Cell Science

369

320

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Cell adhesion, spreading and migration require the dynamic formation and dispersal of contacts with the extracellular matrix (ECM). In vivo, the number, availability and distribution of ECM binding sites dictate the shape of a cell and determine its mobility. To analyse the geometrical limits of ECM binding sites required for cell attachment and spreading, we used microcontact printing to produce regular patterns of ECM protein dots of defined size separated by nonadhesive regions. Cells cultured on these substrata adhere to and spread on ECM regions as small as 0.1 microm2, when spacing between dots is less than 5 microm. Spacing of 5-25 microm induces a cell to adapt its shape to the ECM pattern. The ability to spread and migrate on dots > or =1 microm2 ceases when the dot separation is > or =30 microm. The extent of cell spreading is directly correlated to the total substratum coverage with ECM-proteins, but irrespective of the geometrical pattern. An optimal spreading extent is reached at a surface coating above 15%. Knowledge of these geometrical limits is essential for an understanding of cell adhesion and migration, and for the design of artificial surfaces that optimally interact with cells in a living tissue

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Section: Discussionmentioning

confidence: 99%

Cell behaviour on micropatterned substrata: limits of extracellular matrix geometry for spreading and adhesion

Lehnert

Wehrle-Haller²,

Dávid

et al. 2004

Journal of Cell Science

369

320

View full text Add to dashboard Cite

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“…Details for fabricating the silicon master, PDMS stamps, and microcontact printing have been reported 28, 29. Figure 1 shows the schematic of the patterning method used here.…”

Section: Methodsmentioning

confidence: 99%

Controlling neurite outgrowth with patterned substrates

Yang

2011

J Biomedical Materials Res

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In vivo, neurons form neurites, one of which develops into the axon while others become dendrites. While this neuritogenesis process is well programmed in vivo, there are limited methods to control the number and location of neurite extension in vitro. Here we report a method to control neuritogenesis by confining neurons in specific regions using cell resistant poly(oligoethyleneglycol methacrylate-co-methacrylic acid (OEGMA-co-MA)) or poly(ethyleneglycol-block-lactic acid) PEG-PLA. Line patterned substrates reduce multiple extension of neurites and stimulate bi-directional neurite budding for PC12 and cortical neurons. PC12 cells on 20 and 30 µm line patterns extended one neurite in each direction along the line pattern while cortical neuron on 20 and 30 µm line patterns extended one or two neurites in each direction along the line pattern. Statistical analysis of neurite lengths revealed that PC12 cells and cortical neurons on line patterns extend longer neurites. The ability to guide formation of neurites on patterned substrates is useful for generating neural networks and promoting neurite elongation.

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“…Excess liquid on the surface was removed by blowing with nitrogen gas, and the surface was stamped for 1 min with a patterned polydimethylsiloxane stamp inked with collagen. The stamp was made as described by LeDuc et al (2002), by using a molding made from the SPR 220-3.0 photoresist (Shipley, Marlboro, MA), patterned with the same photomask as for the SU-8 substrates. The stamp was soaked in 100 g/ml type I collagen (USB, Cleveland, OH) for 45 min and rinsed with PBS briefly.…”

Section: Preparation Of Micropatterned Substrates and Local Mechanicamentioning

confidence: 99%

Retrograde Fluxes of Focal Adhesion Proteins in Response to Cell Migration and Mechanical Signals

Guo

Wang

2007

MBoC

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Recent studies suggest that mechanical signals mediated by the extracellular matrix play an essential role in various physiological and pathological processes; yet, how cells respond to mechanical stimuli remains elusive. Using live cell fluorescence imaging, we found that actin filaments, in association with a number of focal adhesion proteins, including zyxin and vasodilator-stimulated phosphoprotein, undergo retrograde fluxes at focal adhesions in the lamella region. This flux is inversely related to cell migration, such that it is amplified in fibroblasts immobilized on micropatterned islands. In addition, the flux is regulated by mechanical signals, including stretching forces applied to flexible substrates and substrate stiffness. Conditions favoring the flux share the common feature of causing large retrograde displacements of the interior actin cytoskeleton relative to the substrate anchorage site, which may function as a switch translating mechanical input into chemical signals, such as tyrosine phosphorylation. In turn, the stimulation of actin flux at focal adhesions may function as part of a feedback mechanism, regulating structural assembly and force production in relation to cell migration and mechanical load. The retrograde transport of associated focal adhesion proteins may play additional roles in delivering signals from focal adhesions to the interior of the cell. INTRODUCTIONRecent studies suggest that mechanosensing is involved in several physiological processes, including embryogenesis (Newman and Comper, 1990;Beloussov et al., 1994) and wound healing (Hinz et al., 2001;Tomasek et al., 2002), and in pathological processes such as fibrosis and carcinogenesis (Paszek et al., 2005). Adherent cells seem capable of both responding to applied mechanical forces (Tzima et al., 2005) and applying contractile forces to probe mechanical properties of the environment (Discher et al., 2005). The downstream responses include changes in migration (Pelham and Wang, 1997;Sheetz et al., 1998;Lo et al., 2000), cell-cell interactions (Guo et al., 2006), proliferation (Wang et al., 2000Nelson et al., 2005), differentiation (Engler et al., 2004(Engler et al., , 2006, and apoptosis . For cultured adherent cells, focal adhesions are thought to mediate mechanosensing through integrin-mediated anchorage to the extracellular matrix (Larsen et al., 2006). The molecular repertoire of focal adhesions includes Ͼ100 adaptor and signaling proteins (Bershadsky et al., 2006), in addition to associated actomyosin bundles that provide mechanical forces for probing the environment (Choquet et al., 1997), inside-out signaling (Chrzanowska-Wodnicka and Burridge, 1996;Schwartz and Ginsberg, 2002), and cell migration (Ridley et al., 2003). However, few details are available concerning how these components work in concert to generate the responses to mechanical signals.Several aspects have emerged recently as the key features of integrin-mediated mechanosensing. First is the increase in cortical organization and contractility in resp...

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Use of micropatterned adhesive surfaces for control of cell behavior

Cited by 33 publications

References 33 publications

Cell behaviour on micropatterned substrata: limits of extracellular matrix geometry for spreading and adhesion

Cell behaviour on micropatterned substrata: limits of extracellular matrix geometry for spreading and adhesion

Controlling neurite outgrowth with patterned substrates

Retrograde Fluxes of Focal Adhesion Proteins in Response to Cell Migration and Mechanical Signals

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