Cell adhesion and focal contact formation on linear RGD molecular gradients: study of non-linear concentration dependence effects

Lagunas, Anna; Comelles, Jordi; Martínez, Elena; Prats‐Alfonso, Elisabet; Acosta, Gerardo; Alberício, Fernando; Samitier, J.

doi:10.1016/j.nano.2011.08.001

Cited by 43 publications

(34 citation statements)

References 21 publications

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“…1C). This observation is consistent with previous studies [47][48]. Lagunas and coworkers showed that a threshold value of 4.0 pmol/cm 2 was sufficient for full cell attachment and spreading [47].…”

Section: Culture Of Hmscs On Rgd-conjugated Poly(cbma) Hydrogelsupporting

confidence: 93%

Modulation of the stemness and osteogenic differentiation of human mesenchymal stem cells by controlling RGD concentrations of poly(carboxybetaine) hydrogel

Chien

Shih

et al. 2014

Biotechnology Journal

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In vitro modulation of the differentiation status of mesenchymal stem cells (MSCs) is important for their application to regenerative medicine. We suggested that the morphology and differentiation states of MSCs could be modulated by controlling the cell affinity of a substrate. The objective of this study was to investigate the effects of surface bio-adhesive signals on self-renewal and osteogenic differentiation of MSCs using a low-fouling platform. Cell-resistant poly(carboxybetaine) hydrogel was conjugated with 5 μM or 5 mM of cell-adhesive arginine-glycine-aspartic acid (RGD) peptides in order to control the cells' affinity to the substrate. Human mesenchymal stem cells (hMSCs) were cultured on the RGD-modified poly(carboxybetaine) hydrogel and then the cells' states of stemness and osteogenic differentiation were evaluated using reverse-transcriptase polymerase chain reaction. The hMSCs formed three-dimensional spheroids on the 5 μM RGD substrate, while cells on the 5 mM RGD substrate exhibited spreading morphology. Furthermore, cells on the 5 μM RGD hydrogel maintained a better stemness phenotype, while the hMSCs on the 5 mM RGD hydrogel proliferated faster and underwent osteogenic differentiation. In conclusion, the stemness of hMSCs was best maintained on a low RGD surface, while osteogenic differentiation of hMSCs was enhanced on a high RGD surface. monolayer culture [13][14], while MSCs with spheroid morphology tend to sustain pluripotent ability during extended culturing [14][15][16]. Cell adhesion to the extracellular matrix (ECM) in a native tissue is initiated by the binding of cell membrane receptors (e.g. integrins) to ECM adhesion proteins such as fibronectin and laminin [17]. Similarly, cell attachment to artificial substrates is usually mediated by surfacebound adhesion proteins. On the other hand, cell-cell attachment is mediated by other types of cell receptors such as cadherins [18]. We suggest that cell morphology during in vitro culture could be determined by the relative affinity between cell-substrate and cell-cell interactions. When the cell-substrate interaction is stronger than the cell-cell interaction, cells attach to and spread on the substrate. Conversely, when the cell-substrate interaction is weaker than the cell-cell interaction, cells tend to form spheroids on the surface [19].We suggested that by controlling the cell affinity of a substrate the morphology and differentiation status of MSCs could be modulated. To this end, surface conjugation of ECM adhesion proteins such as fibronectin, collagen, and laminin, is a common strategy. Alternatively, conjugation of peptides containing cell-binding domains of ECM adhesion proteins is another popular approach. The advantages of peptides compared to intact proteins include cost effectiveness and reduced vulnerability to denaturation. Among the peptides that mediate cell adhesion, arginine-glycine-aspartic acid (RGD), as found in fibronectin and laminin, is the most commonly applied cell-binding peptide for substrate modification in ...

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Section: Culture Of Hmscs On Rgd-conjugated Poly(cbma) Hydrogelsupporting

confidence: 93%

Modulation of the stemness and osteogenic differentiation of human mesenchymal stem cells by controlling RGD concentrations of poly(carboxybetaine) hydrogel

Chien

Shih

et al. 2014

Biotechnology Journal

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“…The type of ligand [9], [19], ligand density [3], [21], [75], [85] and ligand spacing [2], [4], [23], [86] regulate adhesion formation and force transmission. To examine the ligand affinity of cells, force transmission on beads functionalized with the ECM constituent FN were compared to cRGDfk coated beads.…”

Section: Discussionmentioning

confidence: 99%

Force Mapping during the Formation and Maturation of Cell Adhesion Sites with Multiple Optical Tweezers

Schwingel

Bastmeyer

2013

PLoS ONE

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Focal contacts act as mechanosensors allowing cells to respond to their biomechanical environment. Force transmission through newly formed contact sites is a highly dynamic process requiring a stable link between the intracellular cytoskeleton and the extracellular environment. To simultaneously investigate cellular traction forces in several individual maturing adhesion sites within the same cell, we established a custom-built multiple trap optical tweezers setup. Beads functionalized with fibronectin or RGD-peptides were placed onto the apical surface of a cell and trapped with a maximum force of 160 pN. Cells form adhesion contacts around the beads as demonstrated by vinculin accumulation and start to apply traction forces after 30 seconds. Force transmission was found to strongly depend on bead size, surface density of integrin ligands and bead location on the cell surface. Highest traction forces were measured for beads positioned on the leading edge. For mouse embryonic fibroblasts, traction forces acting on single beads are in the range of 80 pN after 5 minutes. If two beads were positioned parallel to the leading edge and with a center-to-center distance less than 10 µm, traction forces acting on single beads were reduced by 40%. This indicates a spatial and temporal coordination of force development in closely related adhesion sites. We also used our setup to compare traction forces, retrograde transport velocities, and migration velocities between two cell lines (mouse melanoma and fibroblasts) and primary chick fibroblasts. We find that maximal force development differs considerably between the three cell types with the primary cells being the strongest. In addition, we observe a linear relation between force and retrograde transport velocity: a high retrograde transport velocity is associated with strong cellular traction forces. In contrast, migration velocity is inversely related to traction forces and retrograde transport velocity.

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“…We hypothesized that the higher concentration of protein adhered to CNTs will facilitate cell adhesion. 31,32 The appropriate functionalization and protein conjugation to CNTs is a challenging step in preparation of aligned CNT substrates. There are two major types of functionalization approaches including covalent and noncovalent reaction processes.…”

Section: Resultsmentioning

confidence: 99%

Alignment of Carbon Nanotubes: An Approach to Modulate Cell Orientation and Asymmetry

et al. 2014

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Stem cells offer a promising tool in tissue engineering strategies, as their differentiated derivatives can be used to reconstruct most biological tissues. These approaches rely on controlling the biophysical cues that tune the ultimate fate of cells. In this context, significant effort has gone to parse out the role of conflicting matrix-elicited signals (e.g., topography and elasticity) in regulation of macroscopic characteristics of cells (e.g., shape and polarity). A critical hurdle, however, lies in our inability to recapitulate the nanoscale spatiotemporal pattern of these signals. The study presented in this manuscript took an initial step to overcome this challenge by developing a carbon nanotube (CNT)-based substrate for nanoresolution control of focal adhesion formation and cell alignment. The utility of this system was studied using human umbilical vascular endothelial cells (HUVECs) and human embryonic stem cells (hESCs) at a single cell level. Our results demonstrated the ability to control cell orientation by merely controlling the alignment of focal adhesions at a nanoscale size. Our long-term vision is to use these nanoengineered substrates to mimic cell orientation in earlier development and explore the role of polarity in asymmetric division and lineage specification of dividing cells.

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Cell adhesion and focal contact formation on linear RGD molecular gradients: study of non-linear concentration dependence effects

Cited by 43 publications

References 21 publications

Modulation of the stemness and osteogenic differentiation of human mesenchymal stem cells by controlling RGD concentrations of poly(carboxybetaine) hydrogel

Modulation of the stemness and osteogenic differentiation of human mesenchymal stem cells by controlling RGD concentrations of poly(carboxybetaine) hydrogel

Force Mapping during the Formation and Maturation of Cell Adhesion Sites with Multiple Optical Tweezers

Alignment of Carbon Nanotubes: An Approach to Modulate Cell Orientation and Asymmetry

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