Spatiotemporal control of cell adhesion on a self-assembled monolayer having a photocleavable protecting group

Nakanishi, Jun; Kikuchi, Yukiko; Takarada, Tohru; Nakayama, Hidekazu; Yamaguchi, Kazuo; Maeda, Mizuo

doi:10.1016/j.aca.2006.04.059

Cited by 74 publications

(71 citation statements)

References 21 publications

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“…The substrate was placed upside down in a glass-bottom dish filled with water and irradiated using an inverted fluorescence microscope (IX81-PAFM, Olympus, Tokyo, Japan) through a stripe-patterned photomask inserted at the field diaphragm, as described previously [24,25]. After irradiation for 10 J cm −2 (e.g.…”

Section: Patterning His-tagged Proteins and Their Immunofluorescence mentioning

confidence: 99%

Dynamic culture substrate that captures a specific extracellular matrix protein in response to light

Nakanishi

Nakayama

Yamaguchi

et al. 2011

Science and Technology of Advanced Materials

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The development of methods for the off-on switching of immobilization or presentation of cell-adhesive peptides and proteins during cell culture is important because such surfaces are useful for the analysis of the dynamic processes of cell adhesion and migration. This paper describes a chemically functionalized gold substrate that captures a genetically tagged extracellular matrix protein in response to light. The substrate was composed of mixed self-assembled monolayers (SAMs) of three disulfide compounds containing (i) a photocleavable poly(ethylene glycol) (PEG), (ii) nitrilotriacetic acid (NTA) and (iii) hepta(ethylene glycol) (EG 7 ). Although the NTA group has an intrinsic high affinity for oligohistidine tag (His-tag) sequences in its Ni 2+ -ion complex, the interaction was suppressed by the steric hindrance of coexisting PEG on the substrate surface. Upon photoirradiation of the substrate to release the PEG chain from the surface, this interaction became possible and hence the protein was captured at the irradiated regions, while keeping the non-specific adsorption of non-His-tagged proteins blocked by the EG 7 underbrush. In this way, we selectively immobilized a His-tagged fibronectin fragment ) to the irradiated regions. In contrast, when bovine serum albumin-a major serum protein-was added as a non-His-tagged protein, the surface did not permit its capture, with or without irradiation. In agreement with these results, cells were selectively attached to the irradiated patterns only when a His-tagged FNIII 7−10 was added to the medium. These results indicate that the present method is useful for studying the cellular behavior on the specific extracellular matrix protein in cell-culturing environments.

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Section: Patterning His-tagged Proteins and Their Immunofluorescence mentioning

confidence: 99%

Dynamic culture substrate that captures a specific extracellular matrix protein in response to light

Nakanishi

Nakayama

Yamaguchi

et al. 2011

Science and Technology of Advanced Materials

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“…Repeated micro-contact printing steps are difficult to perform, as they require to align all printing steps. Methods in which the substrate is held onto the motorised stage of the microscope are more convenient for the repetition of sequential exposure-grafting steps [see sequential photopatterning in panel E: fibrinogen (green), vitronectin (red) and fibronectin (blue); published with permission from Doyle et al (Doyle et al, 2009)] (Kim et al, 2010;Nakanishi et al, 2006). In addition, they can be performed in the presence of cells and thus can be used to micropattern multiple cell types using simple successive light exposures (Kikuchi et al, 2008b).…”

mentioning

confidence: 99%

“…However, these local electrical desorption requires sophisticated methodologies that use microfabricated electrodes and/or micropatterning of electrosensitive surface coatings (Raghavan et al, 2010a). Photo-patterning methods (see Box 1) allow 'on the fly' micropatterning during cell culture to guide cell movement [see panel B; still images from a time-lapse movie of cell migration towards a photo-activated region, published with permission from Nakanishi et al (Nakanishi et al, 2006)] (Kikuchi et al, 2008a) or to sequentially add new cell types (Kikuchi et al, 2008b). …”

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confidence: 99%

Micropatterning as a tool to decipher cell morphogenesis and functions

Théry

2010

Journal of Cell Science

646

560

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Cell architectureThe physiological cell microenvironment consists of extracellular matrix (ECM) fibres, adjacent cells and extracellular fluids, and the cell adhesion machinery is the first cellular component to encounter it. Upon binding of extracellular ligands, localised signalling is induced with subsequent assembly of the cytoskeleton. These localised events will affect the entire cell architecture, because the intracellular space is physically connected and SummaryIn situ, cells are highly sensitive to geometrical and mechanical constraints from their microenvironment. These parameters are, however, uncontrolled under classic culture conditions, which are thus highly artefactual. Micro-engineering techniques provide tools to modify the chemical properties of cell culture substrates at sub-cellular scales. These can be used to restrict the location and shape of the substrate regions, in which cells can attach, so-called micropatterns. Recent progress in micropatterning techniques has enabled the control of most of the crucial parameters of the cell microenvironment. Engineered micropatterns can provide a micrometer-scale, soft, 3-dimensional, complex and dynamic microenvironment for individual cells or for multi-cellular arrangements. Although artificial, micropatterned substrates allow the reconstitution of physiological in situ conditions for controlled in vitro cell culture and have been used to reveal fundamental cell morphogenetic processes as highlighted in this review. By manipulating micropattern shapes, cells were shown to precisely adapt their cytoskeleton architecture to the geometry of their microenvironment. Remodelling of actin and microtubule networks participates in the adaptation of the entire cell polarity with respect to external constraints. These modifications further impact cell migration, growth and differentiation. Journal of Cell Sciencemechanically supported by a dynamic equilibrium (Ingber, 2003; Ingber, 2006). Integrin-based cell adhesionIntegrins are transmembrane receptors that bind to ECM proteins and to intracellular actin filaments. When cells contact the ECM, they change their shape and spread in a multi-step process that includes cell attachment, formation of membrane protrusion, extension of cell membrane, and formation and contraction of stress fibers, which further stimulate cell attachment, membrane protrusion and cell shape extension.The attachment of the actin cytoskeleton to cell adhesions requires integrin clustering. Nanopatterning methods have led to determine the maximum distance of 60 nm between integrin molecules -a distance that still allows intracellular recruitment of actin filaments and signalling molecules (Arnold et al., 2004). Arrays of adhesive dots have been used to study the formation of filopodia and subsequent spreading steps. Depending on the cell type and the level of Rac activation, cells need a minimal distance between adhesion sites, so that filopodia can bridge them and promote cell spreading (Guillou et al., 2008;Lehnert et al., 2004). Th...

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“…2a). 19,67,68 "Caged" means "protected by a photocleavable protecting group". We have termed this dynamic substrate as a caged culture substrate.…”

Section: ·3 Substrates Responsive To Lightmentioning

confidence: 99%

“…67 In a similar way, we were able to induce cell migration by irradiating a region alongside the patterned cells. 68 We applied this method for the selective induction of cell protrusions led by lamellipodia or filopodia along wide and narrow paths, respectively (Fig. 3d).…”

Section: ·3 Substrates Responsive To Lightmentioning

confidence: 99%

Recent Advances in Cell Micropatterning Techniques for Bioanalytical and Biomedical Sciences

et al. 2008

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Cell micropatterning is a method for controlling the placement of living cells on a substrate surface. [1][2][3][4][5][6][7][8] It is important for a wide range of applications, such as tissue engineering, cellbased drug screening, and fundamental cell biology studies. Most cell micropatterning methods fall into three categories based on strategy: (1) seeding cells on a chemically patterned surface of different cell adhesiveness, (2) seeding cells on a topographically patterned surface, or (3) directed delivery of cells onto discrete regions of a substrate. Although the latter two categories are also important, this review mainly focuses on the first category. This is because it is the most common strategy and it provides tools to study fundamental aspects of cell adhesion at the single protein/receptor molecule level. 9There are excellent reviews that also cover the latter two categories. 3,8,10 We first introduce some of the important applications of cell micropatterning in general. We then describe methods for micropatterning the cell adhesiveness, referring to materials that promote or inhibit cell adhesion. We then move on to much more sophisticated substrates, so-called "dynamic substrates", whose cell adhesiveness can be changed by an external stimulus, such as heat, voltage and light. As an example of dynamic substrates, we describe a caged culture substrate developed by our group. This type of functional substrate opens up new possibilities in bioanalytical and biomedical sciences. Cell micropatterning is an important technique for a wide range of applications, such as tissue engineering, cell-based drug screening, and fundamental cell biology studies. This paper overviews cell patterning techniques based on chemically modified substrates with different degrees of cell adhesiveness. In particular, the focus is on dynamic substrates that change their cell adhesiveness in response to external stimuli, such as heat, voltage, and light. Such substrates allow researchers to achieve an in situ alteration of patterns of cell adhesiveness, which is useful for coculturing multiple cell types and analyzing dynamic cellular activities. As an example of dynamic substrates, we introduce a dynamic substrate based on a caged compound, where we accomplished a light-driven alteration of cell adhesiveness and the analysis of a single cell's motility.

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Spatiotemporal control of cell adhesion on a self-assembled monolayer having a photocleavable protecting group

Cited by 74 publications

References 21 publications

Dynamic culture substrate that captures a specific extracellular matrix protein in response to light

Dynamic culture substrate that captures a specific extracellular matrix protein in response to light

Micropatterning as a tool to decipher cell morphogenesis and functions

Recent Advances in Cell Micropatterning Techniques for Bioanalytical and Biomedical Sciences

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