Patterning N‐type and S‐type neuroblastoma cells with Pluronic F108 and ECM proteins

Corey, Joseph M.; Gertz, Caitlyn C.; Sutton, Thomas J.; Chen, Qiaoran; Mycek, Katherine B.; Wang, Bor Shuen; Martin, Abbey A.; Johnson, Sara; Feldman, Eva L.

doi:10.1002/jbm.a.32485

Cited by 29 publications

(30 citation statements)

References 60 publications

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“…In some cases, as shown in Supplementary Figure S2a, edge sharpness is compromised, possibly because of the yield of some fabricated through-holes, and the resulting lack of appropriate control over diffusion and spreading of ECM protein and F108 copolymer. However, the slight equivocality in sharpness is comparable to those of others 12,22,23 and does not affect the presented results in any way.…”

Section: Configurable 2d Cell Patterningsupporting

confidence: 82%

Configurable 2D and 3D spheroid tissue cultures on bioengineered surfaces with acquisition of epithelial–mesenchymal transition characteristics

et al. 2012

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Both two-dimensional (2D) and three-dimensional (3D) biomaterial-based culture platforms that are capable of mimicking the in vivo microenvironment to recapitulate the physiological conditions are vital tools in a wide range of cellular and clinical research. Here we report tissue cultures in a microfluidic chip that allows deterministic patterning of cells in 2D/3D. The chip contains a cell-supporting membrane bioengineered to attain either 2D or 3D cell patterns by selectable deposition of extracellular matrix molecules. Results show a cell-trapping rate as high as 97% in our microchip. Tuning of the surface enables not only highly controlled geometry of the monolayer (2D) cell mass but also 3D culture of uniformly sized multicellular spheroids. The 3D spheroid culture of human epithelial ovarian cancer cells in the microfluidic chip resulted in acquisition of mesenchymal traits-increased expressions of N-cadherin, vimentin and fibronectin-and lowered expression of epithelial marker (CD326/epithelial cell adhesion molecule) compared with that in traditional 2D cultures, which is indicative of epithelial-mesenchymal transition in the spheres. In conclusion, these results offer new opportunities to achieve active control of 2D cellular patterns and 3D multicellular spheroids on demand, and may be amenable toward the study of the metastatic processes by in vitro modeling.

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Section: Configurable 2d Cell Patterningsupporting

confidence: 82%

Configurable 2D and 3D spheroid tissue cultures on bioengineered surfaces with acquisition of epithelial–mesenchymal transition characteristics

et al. 2012

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“…MSCs incubated with Pluronic F-108 behaved similarly to unlabeled MSCs, while GNT-labeled MSCs showed a decreased response to both fibronectin and collagen I. This observation that Pluronic F-108 does not inhibit cell adhesion while in the presence of ECM proteins has similarly been evidenced with neuroblastoma cells [38]. Additional studies are currently underway to determine whether the GNTs are physically blocking cell adhesion receptor sites or chemically altering the cell adhesion properties of the labeled MSCs.…”

Section: Resultsmentioning

confidence: 89%

Gadonanotubes as magnetic nanolabels for stem cell detection

et al. 2010

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Stem-cell based therapies have emerged as a promising approach in regenerative medicine. In the development of such therapies, the demand for imaging technologies that permit the noninvasive monitoring of transplanted stem cells in vivo is growing. Here, we report the performance of gadolinium-containing carbon nanocapsules, or gadonanotubes (GNTs), as a new T 1 -weighted magnetic resonance imaging (MRI) intracellular labeling agent for pig bone marrow mesenchymal stem cells (MSCs). Without the use of a transfection agent, micromolar concentrations of GNTs can deliver up to 10 9 Gd 3+ ions per cell without compromising cell viability, differentiation potential, proliferation pattern, and phenotype. Imaging 10×10 6 GNT-labeled MSCs demonstrates a nearly two-fold reduction in T 1 relaxation time when compared to unlabeled MSCs at 1.5 T in a clinical MRI scanner, which easily permits the discrimination of GNT-labeled MSCs in a T 1 -weighted MR image. It is anticipated that GNTs will allow in vivo tracking of GNT-labeled MSCs, as well as other mammalian cell types, by T 1 -weighted imaging with greater efficacy than other current technologies now allow.

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“…(26, 27) Pluronic solution was added to each well, incubated for 24 hrs at room temperature, and washed out with PBS. Then each well was loaded with 50 μl of 5.0% (w/w) aqueous PEG phase prepared with growth medium.…”

Section: Methodsmentioning

confidence: 99%

Optimization of Aqueous Biphasic Tumor Spheroid Microtechnology for Anti-cancer Drug Testing in 3D Culture

et al. 2014

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Tumor spheroids are three-dimensional clusters of cancer cells that exhibit characteristics of poorly perfused tumors and hence present a relevant model for testing the efficacy of anti-cancer compounds. The use of spheroids for drug screening is hindered by technological complexities for high throughput generation of consistent size spheroids individually addressable by drug compounds. Here we present and optimize a simple spheroid technology based on the use of an aqueous two-phase system. Cancer cells confined in a drop of the denser aqueous dextran phase are robotically dispensed into a microwell containing the immersion aqueous polyethylene glycol phase. Cells remain within the drop and form a viable spheroid, without a need for any external stimuli. The size of resulting spheroids is sensitive to volume variations of dispensed drops from the air displacement pipetting head of a commercial liquid handling robot. Therefore, we parametrically optimize the process of dispensing of dextran phase drops. For a given cell density, this optimization reproducibly generates consistent size spheroids in standard 96-well plates. In addition, we evaluate the use of a commercial biochemical assay to examine cellular viability of cancer cell spheroids. Spheroids show a dose-dependent response to cisplatin similar to a monolayer culture. However unlike their two-dimensional counterpart, spheroids exhibit resistance to paclitaxel treatment. This technology, which uses only commercially-available reagents and equipment, can potentially expedite anti-cancer drug discovery. Although the use of robotics makes the ATPS spheroid technology particularly useful for drug screening applications, this approach is compatible with simpler liquid handling techniques such as manual micropipetting and offers a straightforward method of 3D cell culture in research laboratories.

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Patterning N‐type and S‐type neuroblastoma cells with Pluronic F108 and ECM proteins

Cited by 29 publications

References 60 publications

Configurable 2D and 3D spheroid tissue cultures on bioengineered surfaces with acquisition of epithelial–mesenchymal transition characteristics

Configurable 2D and 3D spheroid tissue cultures on bioengineered surfaces with acquisition of epithelial–mesenchymal transition characteristics

Gadonanotubes as magnetic nanolabels for stem cell detection

Optimization of Aqueous Biphasic Tumor Spheroid Microtechnology for Anti-cancer Drug Testing in 3D Culture

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