Molding of Hydrogel Microstructures to Create Multiphenotype Cell Microarrays

Koh, Won Gun; Itle, Laura J.; Pishko, Michael V.

doi:10.1021/ac034773s

Cited by 129 publications

(115 citation statements)

References 41 publications

Supporting

Mentioning

114

Contrasting

Order By: Relevance

“…Previous studies in 2-D and 3-D culture work have used photolithographic techniques [31,[33][34][35], laminar flow [36,[49][50][51][52], and combinations thereof [32,53,54], to generate microarrays of photo-reactive polymers. Although these previous studies provide valuable insights into the effects of ECM signals on cell behavior, photolithographic techniques are typically limited to the use of photo-reactive polymers, and laminar flow is dimensionally limited to micro-scale materials.…”

Section: Discussionmentioning

confidence: 99%

“…Photolithographic methods have been used to generate spatially patterned hydrogel structures, in which distinct regions contain specific cell types [31], cell adhesion ligands [32] or ECM chemistries [33,34]. For example, Pishko and coworkers have used photopolymerization within channels [35] or spots [32] to generate PEG microstructures, and demonstrated that multiple mammalian cell types remain viable in spatially patterned hydrogels for up to 7 days. Other recent studies have used microfluidic systems to generate cell-laden peptide-based hydrogels [36].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

An adaptable hydrogel array format for 3-dimensional cell culture and analysis

et al. 2008

View full text Add to dashboard Cite

Hydrogels have been commonly used as model systems for 3-dimensional (3-D) cell biology, as they have material properties that resemble natural extracellular matrices (ECMs), and their cellinteractive properties can be readily adapted in order to address a particular hypothesis. Natural and synthetic hydrogels have been used to gain fundamental insights into virtually all aspects of cell behavior, including cell adhesion, migration, and differentiated function. However, cell responses to complex 3-D environments are difficult to adequately explore due to the large number of variables that must be controlled simultaneously. Here we describe an adaptable, automated approach for 3-D cell culture within hydrogel arrays. Our initial results demonstrate that the hydrogel network chemistry (both natural and synthetic), cell type, cell density, cell adhesion ligand density, and degradability within each array spot can be systematically varied to screen for environments that promote cell viability in a 3-D context. In a testbed application we then demonstrate that a hydrogel array format can be used to identify environments that promote viability of HL-1 cardiomyocytes, a cell line that has not been cultured previously in 3-D hydrogel matrices. Results demonstrate that the fibronectin-derived cell adhesion ligand RGDSP improves HL-1 viability in a dose-dependent manner, and that the effect of RGDSP is particularly pronounced in degrading hydrogel arrays. Importantly, in the presence of 70µM RGDSP, HL-1 cardiomyocyte viability does not decrease even after 7 days of culture in PEG hydrogels. Taken together, our results indicate that the adaptable, array-based format developed in this study may be useful as an enhanced throughput platform for 3-D culture of a variety of cell types.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

An adaptable hydrogel array format for 3-dimensional cell culture and analysis

et al. 2008

View full text Add to dashboard Cite

show abstract

“…Such advances allow the introduction of spatially specific cues in hydrogels, making multicellular constructs, either through co-cultures or multilineage differentiation, a possibility [16][17][18] . Spatial patterning of hydrogels also provides an additional tool for the development of high-throughput screening technology for the rapid investigation of cell-material interactions [19][20][21][22] .…”

Section: Static Hydrogels That Mimic Biophysical Cuesmentioning

confidence: 99%

Moving from static to dynamic complexity in hydrogel design

2012

View full text Add to dashboard Cite

Hydrogels are water-swollen polymer networks that have found a range of applications from biological scaffolds to contact lenses. Historically, their design has consisted primarily of static systems and those that exhibit simple degradation. However, advances in polymer synthesis and processing have led to a new generation of dynamic systems that are capable of responding to artificial triggers and biological signals with spatial precision. These systems will open up new possibilities for the use of hydrogels as model biological structures and in tissue regeneration.H ydrogels are water-swollen polymer networks that have been used for many decades, with applications as varied as contact lenses and super-absorbant materials. As the field of biomedical engineering has developed, hydrogels have become a prime candidate for application as molecule delivery vehicles and as carriers for cells in tissue engineering, owing to their ability to mimic many aspects of the native cellular environment (for example, high water content, mechanical properties that match soft tissues). Traditional hydrogels, formed through the covalent and non-covalent crosslinking of polymer chains, were regarded as relatively inert materials, providing a simple biomimetic three-dimensional (3D) environment, either for tissue production by local resident cells or for positioning of cells delivered in vivo. However, the simplicity of these materials may have in fact hindered their application, restricting cellular interactions with the environment and preventing uniform extracellular matrix (ECM) production and proper tissue development. In addition, these materials were limited to modelling static environments and lacked the spatiotemporal dynamic properties relevant for complex tissue processes. Fortunately, during the last decade the concepts of hydrogel design and cellular interaction have evolved, shedding light on how they may control cell behaviour, particularly for tissue engineering applications.Hydrogels with a range of mechanical properties, and capable of incorporating a wide range of biologically relevant molecules, from individual functional groups to multidomain proteins, are currently in development 1 . In addition, hydrogels are being designed with spatial heterogeneity, to either replicate properties in native tissue structures or to produce constructs with distinct regionally specific cell behavior 2 . As a result, studies to date have clearly demonstrated the possibility of creating well-defined microenvironments with control over the 3D presentation of signals to cells 3 . However, recently there has been a focus on the concept of hydrogels that exhibit dynamic complexity. These materials should evolve with time and in response to

show abstract

“…Although it is possible to UV crosslink PEG pre-polymer inside microchannels, 11,17,18 the direct crosslinking of the PEG polymer within microfluidic channels has not been shown to generate features with both exposed and non-exposed substrates. Therefore, we hypothesized that the molding of the PDMS stamp on a polymer film would allow for more control over the features of the microfluidic channel.…”

Section: Fabrication Of the Devicementioning

confidence: 99%

Molded polyethylene glycol microstructures for capturing cells within microfluidic channels

et al. 2004

View full text Add to dashboard Cite

The ability to control the deposition and location of adherent and non-adherent cells within microfluidic devices is beneficial for the development of micro-scale bioanalytical tools and high-throughput screening systems. Here, we introduce a simple technique to fabricate poly(ethylene glycol) (PEG) microstructures within microfluidic channels that can be used to dock cells within pre-defined locations. Microstructures of various shapes were used to capture and shear-protect cells despite medium flow in the channel. Using this approach, PEG microwells were fabricated either with exposed or non-exposed substrates. Proteins and cells adhered within microwells with exposed substrates, while non-exposed substrates prevented protein and cell adhesion (although the cells were captured inside the features). Furthermore, immobilized cells remained viable and were stained for cell surface receptors by sequential flow of antibodies and secondary fluorescent probes. With its unique strengths in utility and control, this approach is potentially beneficial for the development of cell-based analytical devices and microreactors that enable the capture and real-time analysis of cells within microchannels, irrespective of cell anchorage properties.

show abstract

Molding of Hydrogel Microstructures to Create Multiphenotype Cell Microarrays

Cited by 129 publications

References 41 publications

An adaptable hydrogel array format for 3-dimensional cell culture and analysis

An adaptable hydrogel array format for 3-dimensional cell culture and analysis

Moving from static to dynamic complexity in hydrogel design

Molded polyethylene glycol microstructures for capturing cells within microfluidic channels

Contact Info

Product

Resources

About