Geometric microenvironment directs cell morphology on topographically patterned hydrogel substrates

Poellmann, Michael J.; Harrell, Patrick A.; King, William P.; Johnson, Amy J. Wagoner

doi:10.1016/j.actbio.2010.03.041

Cited by 45 publications

(39 citation statements)

References 43 publications

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“…extracellular matrix (ECM), self-assembled monolayer (SAM) to print on substrates. The precisely defined ECM micro-patterns allow to study the effect of geometric cues on various important cell functionalities including cell adhesion and spreading 19, 22, 23 , cell proliferation and differentiation 1, 24 , cell fate switching between apoptosis and growth 2 , cell polarity 5, 25 , migration 16, 26–28 , cellular internal compartment organization 16, 25, 29 and the orientation of the axis of division 30, 31 . Increasing evidence suggests that appropriate substrate stiffness can give rise to specific cellular responses 9, 32–40 .…”

Section: Introductionmentioning

confidence: 99%

A novel technique for micro-patterning proteins and cells on polyacrylamide gels

2012

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Spatial patterning of proteins (extracellular matrix, ECM) for living cells on polyacrylamide (PA) hydrogels has been technically challenging due to the compliant nature of the hydrogels and their aqueous environment. Traditional micro-fabrication process is not applicable. Here we report a simple, novel and general method to pattern a variety of commonly used cell adhesion molecules, i.e. Fibronectin (FN), Laminin (LN) and Collagen I (CN), etc. on PA gels. The pattern is first printed on a hydrophilic glass using polydimethylsiloxane (PDMS) stamp and micro-contact printing (μCP). Pre-polymerization solution is applied on the patterned glass and is then sandwiched by a functionalized glass slide, which covalently binds to the gel. The hydrophilic glass slide is then peeled off from the gel when the protein patterns detach from the glass, but remain intact with the gel. The pattern is thus transferred to the gel. The mechanism of pattern transfer is studied in light of interfacial mechanics. It is found that hydrophilic glass offers strong enough adhesion with ECM proteins such that a pattern can be printed, but weak enough adhesion such that they can be completely peeled off by the polymerized gel. This balance is essential for successful pattern transfer. As a demonstration, lines of FN, LN and CN with widths varying from 5–400 μm are patterned on PA gels. Normal fibroblasts (MKF) are cultured on the gel surfaces. The cell attachment and proliferation are confined within these patterns. The method avoids the use of any toxic chemistry often used to pattern different proteins on gel surfaces.

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Section: Introductionmentioning

confidence: 99%

A novel technique for micro-patterning proteins and cells on polyacrylamide gels

2012

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“…Microwells on polyethylene glycol (PEG) could also modulate differentiation of adipocytes or neurons in MSCs [25]. Meanwhile, grid or lattice topography on PA and PDMS substrates are associated with murine MSC morphology and adhesion [26,27], whereas pillar and hexagonal substrates are related with drug or gene delivery to hMSCs [28,29]. Recently, 2176 randomized libraries of surface topographies designed from mathematical algorithms were applied to map the interplay between cells and surface topography and to unravel the unique, formerly unknown, topographies that can induce MSC proliferation or osteogenic differentiation [30].…”

Section: Introductionmentioning

confidence: 99%

Differential regulation of stiffness, topography, and dimension of substrates in rat mesenchymal stem cells

et al. 2013

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a b s t r a c tThe physiological microenvironment of the stem cell niche, including the three factors of stiffness, topography, and dimension, is crucial to stem cell proliferation and differentiation. Although a growing body of evidence is present to elucidate the importance of these factors individually, the interaction of the biophysical parameters of the factors remains insufficiently characterized, particularly for stem cells. To address this issue fully, we applied a micro-fabricated polyacrylamide hydrogel substrate with two elasticities, two topographies, and three dimensions to systematically test proliferation, morphology and spreading, differentiation, and cytoskeletal re-organization of rat bone marrow mesenchymal stem cells (rBMSCs) on twelve cases. An isolated but not combinatory impact of the factors was found regarding the specific functions. Substrate stiffness or dimension is predominant in regulating cell proliferation by fostering cell growth on stiff, unevenly dimensioned substrate. Topography is a key factor for manipulating cell morphology and spreading via the formation of a large spherical shape in a pillar substrate but not in a grooved substrate. Although stiffness leads to osteogenic or neuronal differentiation of rBMSCs on a stiff or soft substrate, respectively, topography or dimension also plays a lesser role in directing cell differentiation. Neither an isolated effect nor a combinatory effect was found for actin or tubulin expression, whereas a seemingly combinatory effect of topography and dimension was found in manipulating vimentin expression. These results further the understandings of stem cell proliferation, morphology, and differentiation in a physiologically mimicking microenvironment.

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“…The structural complexity experienced by cells migrating in an in vivo microenvironment has been recreated in vitro using complex protein patterns [20-24] and complex physical topographies [25,26]. These features approximate the complexity of an in vivo environment, but the analysis of migration on patterned and textured substrates has been restricted to single cell migration because these delicate substrates are disrupted in scratch assays.…”

Section: Resultsmentioning

confidence: 99%

Magnetically attachable stencils and the non-destructive analysis of the contribution made by the underlying matrix to cell migration

Ashby

Wikswo²,

Zijlstra

2012

Biomaterials

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Cell migration is controlled by the integration of numerous distinct components. Consequently, the analysis of cell migration is advancing towards comprehensive, multifaceted in vitro models. To accurately evaluate the contribution of an underlying substrate to cell motility in complex cellular environments we developed a migration assay using magnetically attachable stencils (MAts). When attached to a culture surface, MAts create a defined void in the cell monolayer without disrupting the cells or damaging the underlying substrate. Quantitative analysis of migration into this void reveals the substrate's contribution to migration. The magnetically-guided placement of a microfabricated stencil allows for full experimental control of the substrate on which migration is analyzed. MAts enable the evaluation of intact, defined matrix, and make it possible to analyze migration on unique surfaces such as micropatterned proteins, nano-textured surfaces, and pliable hydrogels. These studies also revealed that mechanical disruption, including the damage that occurs during scratch assays, diminishes migration and confounds the analysis of individual cell behavior. Analysis of migration on increasingly complex biomaterials reveals that the contribution of the underlying matrix depends not only on its molecular composition but also its organization and the context in which it is presented.

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Geometric microenvironment directs cell morphology on topographically patterned hydrogel substrates

Cited by 45 publications

References 43 publications

A novel technique for micro-patterning proteins and cells on polyacrylamide gels

A novel technique for micro-patterning proteins and cells on polyacrylamide gels

Differential regulation of stiffness, topography, and dimension of substrates in rat mesenchymal stem cells

Magnetically attachable stencils and the non-destructive analysis of the contribution made by the underlying matrix to cell migration

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