Stable Non‐Covalent Large Area Patterning of Inert Teflon‐AF Surface: A New Approach to Multiscale Cell Guidance

Valle, Francesco; Chelli, Beatrice; Bianchi, Michele; Greco, Pierpaolo; Bystrenová, Eva; Tonazzini, Ilaria; Biscarini, Fabio

doi:10.1002/adem.201080022

Cited by 22 publications

(25 citation statements)

References 33 publications

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“…Owing to the recent developments in micro/nanoengineering techniques, the processes that control neuronal guidance and differentiation can now be directly investigated using nano/microtextured substrates [23][24][25][26][27][28]. In particular, in the last few years nanogratings-anisotropic topographies composed of alternating lines of grooves and ridges with submicrometre lateral dimensions-have been intensively investigated and have emerged as one of the most effective systems for inducing neuronal alignment via pure cell mechanotransduction.…”

Section: Introductionmentioning

confidence: 99%

Interaction of leech neurons with topographical gratings: comparison with rodent and human neuronal lines and primary cells

et al. 2014

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Controlling and improving neuronal cell migration and neurite outgrowth are critical elements of tissue engineering applications and development of artificial neuronal interfaces. To this end, a promising approach exploits nano/microstructured surfaces, which have been demonstrated to be capable of tuning neuronal differentiation, polarity, migration and neurite orientation. Here, we investigate the neurite contact guidance of leech neurons on plastic gratings (GRs; anisotropic topographies composed of alternating lines of grooves and ridges). By high-resolution microscopy, we quantitatively evaluate the changes in tubulin cytoskeleton organization and cell morphology and in the neurite and growth cone development. The topography-reading process of leech neurons on GRs is mediated by filopodia and is more responsive to 4-µm-period GRs than to smaller period GRs. Leech neuron behaviour on GRs is finally compared and validated with several other neuronal cells, from murine differentiated embryonic stem cells and primary hippocampal neurons to differentiated human neuroblastoma cells.

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Section: Introductionmentioning

confidence: 99%

Interaction of leech neurons with topographical gratings: comparison with rodent and human neuronal lines and primary cells

et al. 2014

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“…We have recently demonstrated that cell guidance may be achieved even on anti-fouling surfaces, such as Teflon, by patterning laminin stripes [42]. In such substrates, the adhesion propensity gradient of the cells between the pattern and the substrate was even sharper.…”

Section: Discussionmentioning

confidence: 99%

“…Poly-(dimethylsiloxane) (PDMS) stamps were prepared by replica moulding of a master fabricated by photolithography as reported in [42]. The PDMS stamp has 22 parallel channels, whose length is 8 mm, and whose width ranges from about 15 to 70 mm, height is 1.5 mm and are spaced apart 160 and 230 mm.…”

Section: Fabrication Of Laminin Patternsmentioning

confidence: 99%

“…These techniques exploit chemical and physical interactions for the deposition/adsorption/self-assembly of the biochemical cues at precise positions on the surface. Linear gradients exhibiting a slope in excess of 0.06 mg(ml mm) 21 of laminin [12] have been developed as a route to fabricate protein-bound surfaces, which, in turn, may induce cell patterning [41][42][43][44]. These methods can be used to create gradients through the interplay between capillary forces and self-organization phenomena occurring in the confined region between the stamp protrusions and the surface [41,45].…”

Section: Introductionmentioning

confidence: 99%

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Neural cell alignment by patterning gradients of the extracellular matrix protein laminin

et al. 2014

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Anisotropic orientation and accurate positioning of neural cells is achieved by patterning stripes of the extracellular matrix protein laminin on the surface of polystyrene tissue culture dishes by micromoulding in capillaries (MIMICs). Laminin concentration decreases from the entrance of the channels in contact with the reservoir towards the end. Immunofluorescence analysis of laminin shows a decreasing gradient of concentration along the longitudinal direction of the stripes. The explanation is the superposition of diffusion and convection of the solute, the former dominating at length scales near the entrance (characteristic length around 50 μm), the latter further away (length scale in excess of 900 μm). These length scales are independent of the channel width explored from about 15 to 45 μm. Neural cells are randomly seeded and selectively adhere to the pattern, leaving the unpatterned areas depleted even upon 6 days of incubation. Cell alignment was assessed by the orientation of the long axis of the 4′,6-diamidino-2-phenylindole-stained nuclei. Samples on patterned the laminin area exhibit a large orientational order parameter. As control, cells on the unpatterned laminin film exhibit no preferential orientation. This implies that the anisotropy of laminin stripes is an effective chemical stimulus for cell recruiting and alignment.

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“…17 Briefly, the Teflon-AF substrates were realized by spin coating glass slides with a solution of Teflon-AF MIMIC was used to fabricate the pattern on the polystyrene (PS) culture dishes (35 mm, Corning, Corning, NY). This technique is based on filling the channels formed by the recesses of an elastomeric stamp placed in contact with a surface with a solution.…”

Section: Methodsmentioning

confidence: 99%

Cell Fluidics: Producing Cellular Streams on Micropatterned Synthetic Surfaces

et al. 2011

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Patterning cell-adhesive molecules on material surfaces provides a powerful tool for controlling and guiding cell locomotion and migration. Here we report fast, reliable, easy to implement methods to fabricate large patterns of proteins on synthetic substrates to control the direction and speed of cells. Two common materials exhibiting very different protein adsorption capacities, namely, polystyrene and Teflon, were functionalized with micrometric stripes of laminin. The protein was noncovalently immobilized onto the surface by following either lithographically controlled wetting (LCW) or micromolding in capillaries (MIMIC). These techniques proved to be sufficiently mild so as not to interfere with the protein adhesion capability. Cells adhered onto the functionalized stripes and remained viable for more than 20 h. During this time frame, cells migrated along the lanes and the dynamics of motion was strongly affected by the substrate surface chemistry and culturing conditions. In particular, enhanced mismatches of cell adhesive properties obtained by the juxtaposition of bare and laminin-functionalized Teflon caused cells to move slowly and their movement to be highly confined. The patterning procedure was also effective at guiding migration on conventional cell culture dishes that were functionalized with laminin patterns, even in the presence of serum proteins, although to a lesser extent compared to that for Teflon. This work demonstrates the possibility of creating well-defined, long-range cellular streams on synthetic substrates by pursuing straightforward functionalizing techniques that can be implemented for a broad class of materials under conventional, long-time cell-culturing conditions. The procedure effectively confines cells to migrate along predefined patterns and can be implemented in different biomedical and biotechnological applications.

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Stable Non‐Covalent Large Area Patterning of Inert Teflon‐AF Surface: A New Approach to Multiscale Cell Guidance

Cited by 22 publications

References 33 publications

Interaction of leech neurons with topographical gratings: comparison with rodent and human neuronal lines and primary cells

Interaction of leech neurons with topographical gratings: comparison with rodent and human neuronal lines and primary cells

Neural cell alignment by patterning gradients of the extracellular matrix protein laminin

Cell Fluidics: Producing Cellular Streams on Micropatterned Synthetic Surfaces

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