Regiospecific Control of Protein Expression in Cells Cultured on Two-Component Counter Gradients of Extracellular Matrix Proteins

Gunawan, Rico C.; Choban, Eric R.; Conour, John E.; Silvestre, Jonathan; Schook, Lawrence B.; Gaskins, H. Rex; Leckband, Deborah; Kenis, Paul J. A.

doi:10.1021/la048303k

Cited by 42 publications

(49 citation statements)

References 23 publications

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“…84 Gunawan and co-workers used gradients of extracellular matrix proteins (laminin and collagen I) created in microfluidic networks to demonstrate control over the expression levels of two proteins linked to cell cycle progression by virtue of the spatial location of cells on the gradients. 156,157 A microfluidic setup was recently used to vary the pH of solution gradually, which enabled a dynamic nonequilibrium study of the self-assembly of collagen molecules. 158 A few exotic types of gradients were created by varying external fields, such as temperature, [159][160][161][162] pH, 99,163 and electrochemical potential.…”

Section: Gradient Temporal Dependencymentioning

confidence: 99%

“…biotin). 10,24,123,156,149,217,226,[229][230][231][232][233][234][235][236][237][238][239][240] In addition, surface-bound gradients have also been used to investigate systematically platelet adhesion, 241,242 enzyme immobilization, 243 cell adhesion ( Figures 15 and 16), 11,21,31,64,65,[72][73][74]108,117,118,139,140,142,150,235,[244][245][246][247][248][249][250][251][252][253][254][255][256][257][258]…”

Section: Screening a Phenomenonmentioning

confidence: 99%

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Surface-Bound Soft Matter Gradients

2008

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This feature article describes the progress realized over the past half century in the field of surface-bound gradient structures created on or from soft materials (oligomers and/or polymers), or those enabling the study of the behavior of soft materials. By highlighting our work in the field and accounting for the contribution of other groups, we emphasize the exceptional versatility of gradient assemblies in facilitating fast screening of physicochemical phenomena, acting as "recording media" for monitoring a process, and playing a key role in the design and fabrication of surface-bound molecular and macromolecular motors capable of directing a transport phenomenon.

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Section: Gradient Temporal Dependencymentioning

confidence: 99%

Section: Screening a Phenomenonmentioning

confidence: 99%

Surface-Bound Soft Matter Gradients

2008

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“…To achieve this, carboxy-terminated beads were activated to bind primary amines of acrylamide through 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC)/N-hydroxy-succinimide ester (NHS) coupling chemistry. 8 Initial tests showed that activated beads added directly to the gelation mixture interfered with the polymerization. This was avoided by first activating beads with EDC/NHS.…”

Section: Microbead Preparationmentioning

confidence: 99%

Measuring Traction Forces in Long-Term Cell Cultures

Mann

Leckband

2010

Cel. Mol. Bioeng.

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This report describes an approach to extend indefinitely the duration of traction force measurements with cells cultured on soft polyacrylamide gels. Typical observation times in traction force measurements on similar substrates have been limited to 24-48 h, but cell differentiation or responses to external stimuli often occur over much longer periods of several days or weeks. This study describes a method for covalently linking fluorescent marker beads to a polyacrylamide matrix that renders the hydrogels useful for traction force measurements over several days. This approach was validated by comparing the contractility of C2C12 murine skeletal muscle cells prior to myotube formation, after one day in culture, with that of myotubes after 7 days in culture. Measured tractions increased concurrent with the differentiation of C2C12 cells to the contractile, myotube phenotype. Covalent bead linkage thus extends the useful period during which traction force data can be obtained with cells cultured on optically transparent polyacrylamide hydrogels with controlled elastic moduli.

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“…[4][5][6][7] However, current microfluidic approaches for cell study focus on culturing cells on glass or plastic substrates with gradients of diffusible proteins or surface-bound biomolecules. [8][9][10][11] In native tissues, cells always interact with the surrounding microenvironment with various biophysical properties. 12,13 So, it is preferred to use more physiological cell culture substrate to better mimic the natural cell microenvironment.…”

Section: Introductionmentioning

confidence: 99%

Covalently immobilized biomolecule gradient on hydrogel surface using a gradient generating microfluidic device for a quantitative mesenchymal stem cell study

Liu

Liang

et al. 2012

Biomicrofluidics

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Precisely controlling the spatial distribution of biomolecules on biomaterial surface is important for directing cellular activities in the controlled cell microenvironment. This paper describes a polydimethylsiloxane (PDMS) gradient-generating microfluidic device to immobilize the gradient of cellular adhesive Arg-Gly-Asp (RGD) peptide on poly (ethylene glycol) (PEG) hydrogel. Hydrogels are formed by exposing the mixture of PEG diacrylate (PEGDA), acryloyl-PEG-RGD, and photo-initiator with ultraviolet light. The microfluidic chip was simulated by a fluid dynamic model for the biomolecule diffusion process and gradient generation. PEG hydrogel covalently immobilized with RGD peptide gradient was fabricated in this microfluidic device by photo-polymerization. Bone marrow derived rat mesenchymal stem cells (MSCs) were then cultured on the surface of RGD gradient PEG hydrogel. Cell adhesion of rat MSCs on PEG hydrogel with various RGD gradients were then qualitatively and quantitatively analyzed by immunostaining method. MSCs cultured on PEG hydrogel surface with RGD gradient showed a grated fashion for cell adhesion and spreading that was proportional to RGD concentration. It was also found that 0.107-0.143 mM was the critical RGD concentration range for MSCs maximum adhesion on PEG hydrogel.

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Regiospecific Control of Protein Expression in Cells Cultured on Two-Component Counter Gradients of Extracellular Matrix Proteins

Cited by 42 publications

References 23 publications

Surface-Bound Soft Matter Gradients

Surface-Bound Soft Matter Gradients

Measuring Traction Forces in Long-Term Cell Cultures

Covalently immobilized biomolecule gradient on hydrogel surface using a gradient generating microfluidic device for a quantitative mesenchymal stem cell study

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