Microcontact Printing of Proteins on Mixed Self-Assembled Monolayers

Tan, John L.; Tien, and Joe; Chen, Christopher

doi:10.1021/la011351+

Cited by 185 publications

(175 citation statements)

References 34 publications

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“…To restrict cell adhesion to the tips of the posts, and thereby precisely define the surface mechanics, we used microcontact printing to deliver fibronectin from a stamp onto the tips of the posts (Fig. 2B), and adsorbed Pluronics F127 onto the remaining unstamped regions of the array to block nonspecific protein adsorption and cell adhesion (31,32). Using this method, we can print fibronectin onto specific posts within the array to spatially pattern the adhesiveness of the surface toward cells (Fig.…”

Section: Resultsmentioning

confidence: 99%

Cells lying on a bed of microneedles: An approach to isolate mechanical force

Tan

Tien

Pirone

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

1,788

1,813

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We describe an approach to manipulate and measure mechanical interactions between cells and their underlying substrates by using microfabricated arrays of elastomeric, microneedle-like posts. By controlling the geometry of the posts, we varied the compliance of the substrate while holding other surface properties constant. Cells attached to, spread across, and deflected multiple posts. The deflections of the posts occurred independently of neighboring posts and, therefore, directly reported the subcellular distribution of traction forces. We report two classes of force-supporting adhesions that exhibit distinct force-size relationships. Force increased with size of adhesions for adhesions larger than 1 m 2 , whereas no such correlation existed for smaller adhesions. By controlling cell adhesion on these micromechanical sensors, we showed that cell morphology regulates the magnitude of traction force generated by cells. Cells that were prevented from spreading and flattening against the substrate did not contract in response to stimulation by serum or lysophosphatidic acid, whereas spread cells did. Contractility in the unspread cells was rescued by expression of constitutively active RhoA. Together, these findings demonstrate a coordination of biochemical and mechanical signals to regulate cell adhesion and mechanics, and they introduce the use of arrays of mechanically isolated sensors to manipulate and measure the mechanical interactions of cells.

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Section: Resultsmentioning

confidence: 99%

Cells lying on a bed of microneedles: An approach to isolate mechanical force

Tan

Tien

Pirone

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

1,788

1,813

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“…The microcontact printing process employed a published technique by Tan (Tan, Tien et al 2002;Tan, Liu et al 2004). Briefly, Poly(dimethylsiloxane) (PDMS) circular stamps with diameter, D (400 µm and 800 µm), and pitch, P (500 µm) were fabricated using standard soft lithography techniques (Younan Xia 1998).…”

Section: Micropatterned Ebsmentioning

confidence: 99%

Geometric Control of Cardiomyogenic Induction in Human Pluripotent Stem Cells

Bauwens

Song

Thavandiran

et al. 2011

Tissue Engineering Part A

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Pluripotent stem cells provide the opportunity to study human cardiogenesis in vitro, and are a renewable source of tissue for drug testing and disease models, including replacement cardiomyocytes that may be a useful treatment for heart failure. Typically, differentiation is initiated by forming spherical cell aggregates wherein an extraembryonic endoderm (ExE) layer develops on the surface. Given that interactions between endoderm and mesoderm influence embryonic cardiogenesis, we examined the impact of human embryonic stem cell (hESC) aggregate size on endoderm and cardiac development. We first demonstrated aggregate size control by micropatterning hESC colonies at defined diameters and transferring the colonies to suspension. The ratio of endoderm (GATA-6) to neural (PAX6) gene and protein expression increased with decreasing colony size. Subsequently, maximum mesoderm and cardiac induction occurred in larger aggregates when initiated with endoderm-biased hESCs (high GATA-6:PAX6), and in smaller aggregates when initiated with neural-biased hESCs (low GATA-6:PAX6). Additionally, incorporating micropatterned aggregates in a stirred suspension bioreactor increased cell yields and contracting aggregate frequency. We next interrogated the relationship between aggregate size and endoderm and cardiac differentiation efficiency in sizecontrolled aggregates, generated using forced aggregation, in defined cardiogenic medium.

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“…20,21 Briefly, lines of photoresist were micropatterned on 30-mm circular glass cover slips using standard photolithography techniques with a photomask made of Ronchi ruling glass (Edmund Industrial Optics). Polydimethylsiloxane (PDMS) (Sylgard 184, Dow Corning) stamps were then cast against the micropatterned glass cover slips overnight at 65°C.…”

Section: Micropatterningmentioning

confidence: 99%

Cardiomyocyte Cultures With Controlled Macroscopic Anisotropy

Bursac

Parker

Iravanian

et al. 2002

Circulation Research

239

162

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Abstract-Structural and functional cardiac anisotropy varies with the development, location, and pathophysiology in the heart. The goal of this study was to design a cell culture model system in which the degree, change in fiber direction, and discontinuity of anisotropy can be controlled over centimeter-size length scales. Neonatal rat ventricular myocytes were cultured on fibronectin on 20-mm diameter circular cover slips. Structure-function relationships were assessed using immunostaining and optical mapping. Cell culture on microabraded cover slips yielded cell elongation and coalignment in the direction of abrasion, and uniform, macroscopically continuous, elliptical propagation with point stimulation. Coarser microabrasion (wider and deeper abrasion grooves) increased longitudinal (23.5 to 37.2 cm/s; rϭ0.66) and decreased transverse conduction velocity (18.1 to 9.2 cm/s; rϭϪ0.84), which resulted in increased longitudinal-to-transverse velocity anisotropy ratios (1.3 to 3.7, nϭ61). A thin transition zone between adjacent uniformly anisotropic areas with 45°or 90°difference in fiber orientation acted as a secondary source during 2ϫ threshold field stimulus. Cell culture on cover slips micropatterned with 12-or 25-m wide fibronectin lines and previously coated with decreasing concentrations of background fibronectin yielded transition from continuous to discontinuous anisotropic architecture with longitudinally oriented intercellular clefts, decreased transverse velocity (16.9 to 2.6 cm/s; rϭϪ0.95), increased velocity anisotropy ratios (1.6 to 5.6, nϭ70), and decreased longitudinal velocity (36.4 to 14.6 cm/s; rϭϪ0.85) for anisotropy ratios Ͼ3.5. Cultures of cardiac myocytes with controlled degree, uniformity and continuity of structural, and functional anisotropy may enable systematic 2-dimensional in vitro studies of macroscopic structure-related mechanisms of reentrant arrhythmias.

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Microcontact Printing of Proteins on Mixed Self-Assembled Monolayers

Cited by 185 publications

References 34 publications

Cells lying on a bed of microneedles: An approach to isolate mechanical force

Cells lying on a bed of microneedles: An approach to isolate mechanical force

Geometric Control of Cardiomyogenic Induction in Human Pluripotent Stem Cells

Cardiomyocyte Cultures With Controlled Macroscopic Anisotropy

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