Engineering subcellular-patterned biointerfaces to regulate the surface wetting of multicellular spheroids

Several works reported increased differentiation of neuronal cells grown on graphene; however, the molecular mechanism driving axon elongation on this material has remained elusive. Here, we study the axonal transport of nerve growth factor (NGF), the neurotrophin supporting development of peripheral neurons, as a key player in the time course of axonal elongation of dorsal root ganglion neurons on graphene. We find that graphene drastically reduces the number of retrogradely transported NGF vesicles in favor of a stalled population in the first two days of culture, in which the boost of axon elongation is observed. This correlates with a mutual charge redistribution, observed via Raman spectroscopy and electrophysiological recordings. Furthermore, ultrastructural analysis indicates a reduced microtubule distance and an elongated axonal topology. Thus, both electrophysiological and structural effects can account for graphene action on neuron development. Unraveling the molecular players underneath this interplay may open new avenues for axon regeneration applications.

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“…Moreover, as graphene crumpling was never observed in our analyses, we can exclude its effect on substrate wettability and soma clumping 21,23 .…”

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confidence: 73%

“…S2). The flatness of the surface, with a root mean square roughness of about 1.8 nm, made us exclude the presence of features that can lead to discontinuous cell-substrate adhesion 21 .…”

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confidence: 99%

Graphene Promotes Axon Elongation through Local Stall of Nerve Growth Factor Signaling Endosomes

et al. 2020

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“…Current well established and routine methods for 3D spheroids fabrication include spinner flasks, bacterial grade dishes, culturing cells on ultralow attachment surfaces including (ULA) plates and method of hanging drop . Using spinner flasks, bacterial grade dishes, and ULA plates result in formation of multiple spheroids per flask/dish/well that show high variability in size.…”

Section: Introductionmentioning

confidence: 99%

Facile One Step Formation and Screening of Tumor Spheroids Using Droplet‐Microarray Platform

Popova

Tronser

Demir

et al. 2019

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Tumor spheroids or microtumors are important 3D in vitro tumor models that closely resemble a tumor's in vivo “microenvironment” compared to 2D cell culture. Microtumors are widely applied in the fields of fundamental cancer research, drug discovery, and precision medicine. In precision medicine tumor spheroids derived from patient tumor cells represent a promising system for drug sensitivity and resistance testing. Established and commonly used platforms for routine screenings of cell spheroids, based on microtiter plates of 96‐ and 384‐well formats, require relatively large numbers of cells and compounds, and often lead to the formation of multiple spheroids per well. In this study, an application of the Droplet Microarray platform, based on hydrophilic–superhydrophobic patterning, in combination with the method of hanging droplet, is demonstrated for the formation of highly miniaturized single‐spheroid‐microarrays. Formation of spheroids from several commonly used cancer cell lines in 100 nL droplets starting with as few as 150 cells per spheroid within 24–48 h is demonstrated. Established methodology carries a potential to be adopted for routine workflows of high‐throughput compound screening in 3D cancer spheroids or microtumors, which is crucial for the fields of fundamental cancer research, drug discovery, and precision medicine.

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“…Generally, it is difficult to study the self‐organization process in living organisms in vivo because it is impractical to change many factors that determine the behavior of living systems. Recently, 3D cellular aggregates, instead of 2Dcell sheets, have been employed as building blocks in order to understand the principles of cell–cell and cell–matrix adhesion, as well as cell sorting, because 3D cellular aggregates can better simulate real cell morphology, fate and metabolic activities in vivo. To organize these cellular aggregates, biodegradable polymers, and hydrogels, have been employed as supporting scaffolds for seeding or encasing cells, including alginate with extracellular matrix (ECM) proteins, poly(ethylene glycol) diacrylate (PEGDA) and methacrylated gelatine (GelMA).…”

Section: Introductionmentioning

confidence: 99%

“…For example, Vrij et al used pure cells as a living, self‐scaffolding building block for the free‐form fabrication of stable tissue constructs with defined complex geometries by sequential self‐organization. By building these 3D cellular aggregates, enormous efforts have been devoted to studying the topographic interactions between cellular aggregates and substrates, or the macroscale wetting and fusion of cellular aggregates . However, it remains unclear how topography affects self‐organization of cellular aggregates.…”

Section: Introductionmentioning

confidence: 99%

Topography‐Induced Cell Self‐Organization from Simple to Complex Aggregates

Luo

Meng

et al. 2019

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Self‐organization is a fundamental and indispensable process in a living system. To understand cell behavior in vivo such as tumorigenesis, 3D cellular aggregates, instead of 2D cellular sheets, have been employed as a vivid in vitro model for self‐organization. However, most focus on the macroscale wetting and fusion of cellular aggregates. In this study, it is reported that self‐organization of cells from simple to complex aggregates can be induced by multiscale topography through confined templates at the macroscale and cell interactions at the nanoscale. On the one hand, macroscale templates are beneficial for the organization of individual cells into simple and complex cellular aggregates with various shapes. On the other hand, the realization of these macro‐organizations also depends on cell interactions at the nanoscale, as demonstrated by the intimate contact between nanoscale pseudopodia stretched by adjacent frontier cells, much like holding hands and by the variation in the intermolecular interactions based on E‐cadherin. Therefore, these findings may be very meaningful for clarifying the organizational mechanism of tumor development, tissue engineering and regenerative medicine.

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Engineering subcellular-patterned biointerfaces to regulate the surface wetting of multicellular spheroids

Cited by 14 publications

References 67 publications

Graphene Promotes Axon Elongation through Local Stall of Nerve Growth Factor Signaling Endosomes

Graphene Promotes Axon Elongation through Local Stall of Nerve Growth Factor Signaling Endosomes

Facile One Step Formation and Screening of Tumor Spheroids Using Droplet‐Microarray Platform

Topography‐Induced Cell Self‐Organization from Simple to Complex Aggregates

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