“…Alternatively, PDMS stamp molds were also fabricated by spin coating SU-8 negative photoresist (MicroChem, Newton, MA) onto a silicon wafer, exposing photoresist to UV through a mask for 10 s, and then developing the photoresist following the manufacturer's instructions. The mask was prepared using a computer drawing program to create a pattern that was printed onto a transparency [24,25], also with 1 mm features. PDMS (5.5 g) was poured onto the photolithographic mold, cured at 60 °C for 1-3 h, and then carefully peeled away from mold.…”
Section: Patterned Sams and Complex Depositionmentioning
confidence: 99%
“…SAMs were explored to examine the relationship between surface chemistry and substrate-mediated transfection. SAMs of alkanethiols on gold can be used in cell culture for periods of days [25] and have been used in many cell culture studies, particularly to understand molecular surface determinants required for adhesion dependent cell growth and proliferation [36][37][38].…”
Section: Transfection On Bulk Sams On Goldmentioning
Gene transfer has many potential applications in basic and applied sciences. In vitro, DNA delivery can be enhanced by increasing the concentration of DNA in the cellular microenvironment through immobilization of DNA to a substrate that supports cell adhesion. Substrate-mediated delivery describes the immobilization of DNA, complexed with cationic lipids or polymers, to a biomaterial or substrate. As surface properties are critical to the efficiency of the surface delivery approach, selfassembled monolayers (SAMs) of alkanethiols on gold were used to correlate surface chemistry of the substrate to binding, release, and transfection of non-specifically immobilized complexes. Surface hydrophobicity and ionization were found to mediate both DNA complex immobilization and transfection, but had no effect on complex release. Additionally, SAMs were used in conjunction with soft lithographic techniques to imprint substrates with specific patterns, resulting in patterned DNA complex deposition and transfection, with transfection efficiencies in the patterns nearing 40%. Controlling the interactions between complexes and substrates, with the potential for patterned delivery, can be used to locally enhance or regulate gene transfer, with applications to tissue engineering scaffolds and transfected cell arrays.
“…Alternatively, PDMS stamp molds were also fabricated by spin coating SU-8 negative photoresist (MicroChem, Newton, MA) onto a silicon wafer, exposing photoresist to UV through a mask for 10 s, and then developing the photoresist following the manufacturer's instructions. The mask was prepared using a computer drawing program to create a pattern that was printed onto a transparency [24,25], also with 1 mm features. PDMS (5.5 g) was poured onto the photolithographic mold, cured at 60 °C for 1-3 h, and then carefully peeled away from mold.…”
Section: Patterned Sams and Complex Depositionmentioning
confidence: 99%
“…SAMs were explored to examine the relationship between surface chemistry and substrate-mediated transfection. SAMs of alkanethiols on gold can be used in cell culture for periods of days [25] and have been used in many cell culture studies, particularly to understand molecular surface determinants required for adhesion dependent cell growth and proliferation [36][37][38].…”
Section: Transfection On Bulk Sams On Goldmentioning
Gene transfer has many potential applications in basic and applied sciences. In vitro, DNA delivery can be enhanced by increasing the concentration of DNA in the cellular microenvironment through immobilization of DNA to a substrate that supports cell adhesion. Substrate-mediated delivery describes the immobilization of DNA, complexed with cationic lipids or polymers, to a biomaterial or substrate. As surface properties are critical to the efficiency of the surface delivery approach, selfassembled monolayers (SAMs) of alkanethiols on gold were used to correlate surface chemistry of the substrate to binding, release, and transfection of non-specifically immobilized complexes. Surface hydrophobicity and ionization were found to mediate both DNA complex immobilization and transfection, but had no effect on complex release. Additionally, SAMs were used in conjunction with soft lithographic techniques to imprint substrates with specific patterns, resulting in patterned DNA complex deposition and transfection, with transfection efficiencies in the patterns nearing 40%. Controlling the interactions between complexes and substrates, with the potential for patterned delivery, can be used to locally enhance or regulate gene transfer, with applications to tissue engineering scaffolds and transfected cell arrays.
“…[1][2][3][4][5][6][7][8][9] Micrometerscale patterning of cells on surfaces has used in tissue engineering, biosensors, formation of neuronal networks, and in investigation of fundamental cell biology questions. The general approach to cell patterning has been based on modifying surface properties followed by selective cell attachment and spreading.…”
This paper describes a simple plasma-based dry etching method that enables patterned cell culture inside microfluidic devices by allowing patterning, fluidic bonding and sterilization steps to be carried out in a single step. This plasma-based dry etching method was used to pattern celladhesive and non-adhesive areas on the glass and polystyrene substrates. The patterned substrate was used for selective attachment and growth of human umbilical vein endothelial cells, MDA-MB-231 human breast cancer cells, NIH 3T3 mouse fibroblasts, and primary rat cortical neurons. Finally, we have successfully combined the dry-patterned substrate with a microfluidic device. Patterned primary rat neurons were maintained for up to 6 days inside the microfluidic devices and the neurons' somas and processes were confined to the cell-adhesive region. The method developed in this work offers a convenient way of micropatterning biomaterials for selective attachment of cells on the substrates, and enables culturing of patterned cells inside microfluidic devices for a number of biological research applications where cells need to be exposed to wellcontrolled fluidic microenvironment.
“…9,25,55,64,66,67,73,75,92 . Technology has been applied to cell biology questions using techniques such as soft lithography 28,40,48,58,104 at a cell monolayer or whole cell size-scale. 15,18,27,28,50,52,59,96,119 .…”
Section: Nano-and Microtechnology To Address the Cytoskeletonmentioning
Abstract-Cells function based on a complex set of interactions that control pathways resulting in ultimate cell fates including proliferation, differentiation, and apoptosis. The interworkings of this immensely dense network of intracellular molecules are influenced by more than random protein and nucleic acid distribution where their interactions culminate in distinct cellular function. By probing the design of these biological systems from an engineering perspective, researchers can gain great insight that will aid in building and utilizing systems that are on this size scale where traditional large-scale rules may fail to apply. The organized interaction and gradient distribution in intracellular space imply a structural architecture that modulates cellular processes by influencing biochemical interactions including transport and binding-reactions. One significant structure that plays a role in this modulation is the cell cytoskeleton. Here, we discuss the cytoskeleton as a central and integrating functional structure in influencing cell processes and we describe technology useful for probing this structure. We explain the nanometer scale science of cytoskeletal structure with respect to intracellular organization, mechanotransduction, cytoskeletal-associated proteins, and motor molecules, as well as nano-and microtechnologies that are applicable for experimental studies of the cytoskeleton. This biological architecture of the cytoskeleton influences molecular, cellular, and physiological processes through structured multimodular and hierarchical principles centered on these functional filaments. Through investigating these organic systems that have evolved over billions of years, understanding in biology, engineering, and nanometer-scaled science will be advanced.
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