Hydrogel Cell Cultures

Cushing, Melinda C.; Anseth, Kristi S.

doi:10.1126/science.1140171

Cited by 473 publications

(350 citation statements)

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“…This difference most likely reflects the fact that unlike Matrigel, the worm gels lack the various proteins and growth factors that are known to promote cellular growth and proliferation. [4] [12,26] Sheet-supported 3D cell culture provides a convenient means of handling and analyzing 3D cell cultures, and thermo-responsive hydrogels provide a convenient vehicle to deliver and embed cells into the sheets. The disulfide-functionalized diblock copolymer hydrogel described herein is expected to be preferable to commercial protein-based gels, particularly for applications where the biological effects of such animal-derived gels are not acceptable, or are too variable.…”

Section: Cellular Densities Of A549-gfp Cells Became Indistinguishablmentioning

confidence: 99%

“…[6,11] Moreover, these components often vary in composition and concentration between batches, which can introduce artifacts in cell biology studies. [4,12] Furthermore, such biopolymers have a limited shelf-life and are relatively expensive. [4] In contrast, synthetic hydrogels based on poly(ethylene glycol), polyacrylamide, poly(N-isopropyl acrylamide), poly(vinyl alcohol) or poly(acrylic acid) [10] have a user-defined composition and provide a cost-effective, reproducible and tunable environment for cell studies.…”

mentioning

confidence: 99%

“…that promote cell growth. [4,12] To determine whether cells embedded in these disulfide-based worm gels could still proliferate, we spotted 1 µL suspensions of A549-GFP cells (3×10 4 cells/zone) into worm gels within zones of the mesh sheets, and observed changes in cell density over the course of 12 days. Optical microscopy studies indicated that the density of A549-GFP cells increased progressively from the first to the fifth day of culture (Figure 3A).…”

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confidence: 99%

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Disulfide-Based Diblock Copolymer Worm Gels: A Wholly-Synthetic Thermoreversible 3D Matrix for Sheet-Based Cultures

et al. 2015

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It is well-known that 3D in vitro cell cultures provide a much better model than 2D cell cultures for understanding the in vivo microenvironment of cells. However, significant technical challenges in handling and analyzing 3D cell cultures remain, which currently limits their widespread application. Herein, we demonstrate the application of wholly synthetic thermoresponsive block copolymer worms in sheet-based 3D cell culture. These worms form a soft, free-standing gel reversibly at 20–37 °C, which can be rapidly converted into a free-flowing dispersion of spheres on cooling to 5 °C. Functionalization of the worms with disulfide groups was found to be essential for ensuring sufficient mechanical stability of these hydrogels to enable long-term cell culture. These disulfide groups are conveniently introduced via statistical copolymerization of a disulfide-based dimethacrylate under conditions that favor intramolecular cyclization and subsequent thiol/disulfide exchange leads to the formation of reversible covalent bonds between adjacent worms within the gel. This new approach enables cells to be embedded within micrometer-thick slabs of gel with good viability, permits cell culture for at least 12 days, and facilitates recovery of viable cells from the gel simply by incubating the culture in buffer at 4 °C (thus, avoiding the enzymatic degradation required for cell harvesting when using commercial protein-based gels, such as Matrigel).

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Section: Cellular Densities Of A549-gfp Cells Became Indistinguishablmentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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Disulfide-Based Diblock Copolymer Worm Gels: A Wholly-Synthetic Thermoreversible 3D Matrix for Sheet-Based Cultures

et al. 2015

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“…For a biomaterial to substitute for the ECM, it needs to recapitulate the principal functions of the natural ECM in orchestrating cell proliferation, migration, differentiation, angiogenesis and invasion. 13,14 Recreating the interwoven set of biochemical and mechanical cues in the cellular microenvironment 2,15 and allowing full experimental versatility in fabrication methods for engineered microenvironments 1,16,17 requires consideration of a variety of design criteria. As summarized in Figure 1, the performance criteria that should be met by a biomimetic ECM substitute should include: (1) experimentally variable composition, (2) experimentally variable compliance, (3) controllable biodegradability in vitro and in vivo, (4) multiple physical forms for in situ crosslinking during cell encapsulation and fabrication, (5) batch to batch consistency, (6) ease of use at physiological temperature and pH, (7) transparency for ease of visualization, (8) compatibility with high throughput screening (HTS) platforms, and (9) translational potential.…”

Section: Introductionmentioning

confidence: 99%

“…[1][2][3][4] Among the naturally-derived biomaterials is an in situ-crosslinkable synthetic mimic of the extracellular matrix (sECM) that is based on chemically-modified hyaluronan (HA). 5 HA is a ubiquitous non-sulfated glycosaminoglycan with critical roles in morphogenesis, 6 and chemically-modified HA derivatives have generated considerable interest as biomaterials for tissue engineering.…”

Section: Introductionmentioning

confidence: 99%

Engineering a clinically-useful matrix for cell therapy

Prestwich

2008

Organogenesis

103

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, biological, engineering, marketing, regulatory, and financial constraints. What is required is a biocompatible material for culture of cells in three-dimensions (3-D) that offers ease of use, experimental flexibility to alter composition and compliance, and a composition that would permit a seamless transition from in vitro to in vivo use. The challenge is to replicate the complexity of the native extracellular matrix (ECM) environment with the minimum number of components necessary to allow cells to rebuild a given tissue. Our approach is to deconstruct the ECM to a few modular components that can be reassembled into biomimetic materials that meet these criteria. These semi-synthetic ECMs (sECMs) employ thiol-modified derivatives of hyaluronic acid (HA) that can form covalently crosslinked, biodegradable hydrogels. These sECMs are "living" biopolymers, meaning that they can be crosslinked in the presence of cells or tissues to enable cell therapy and tissue engineering. Moreover, the sECMs allow inclusion of the appropriate biological cues needed to simulate the complexity of the ECM of a given tissue. Taken together, the sECM technology offers a manufacturable, highly reproducible, flexible, FDA-approvable, and affordable vehicle for cell expansion and differentiation in 3-D.

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