Microstructured platforms to study nanotube-mediated long-distance cell-to-cell connections

Abstract

Recently, numerous innovative approaches have attempted to overcome the shortcomings of standard tissue culturing by providing custom-tailored substrates with superior features. In particular, tunable surface chemistry and topographical micro-and nanostructuring have been highlighted as potent effectors to control cell behavior. Apart from tissue engineering and the development of biosensors and diagnostic assays, the need for custom-tailored platform systems is accentuated by a variety of complex and poorly characterized biological processes. One of these processes is cell-to-cell communication mediated by tunneling nanotubes ͑TNTs͒, the reliable statistical analysis of which is consistently hampered by critical dependencies on various experimental factors, such as cell singularization, spacing, and alignment. Here, the authors developed a microstructured platform based on a combination of controlled surface chemistry along with topographic parameters, which permits the controllable attachment of different cell types to complementary patterns of cell attracting/nonattracting surface domains and-as a consequencerepresents a standardized analysis tool to approach a wide range of biological questions. Apart from the technical complementation of mainstream applications, the developed surfaces could successfully be used to statistically determine TNT-based intercellular connection processes as they are occurring in standard as well as primary cell cultures.

Nanotube Action between Human Mesothelial Cells Reveals Novel Aspects of Inflammatory Responses

“…1C). Both findings are in consistency with previous observations made for various other cell types [5], [11], [12]. Frequently, discrete filopodia-like protrusions in contact with the substratum and seemingly directed towards adjacent cells, emitted by individual HPMCs were observable (Fig.…”

Dynamic monitoring of membrane nanotubes formation induced by vaccinia virus on a high throughput microfluidic chip

“…A microgrooved substrate to guide cell alignment has been fabricated to mimic the extracellular matrix topography and explore the virus cell-to-cell spread behavior21. Single PC12 cells have been patterned orderly on biofunctionalized gold microstructures to analyze the influence of cell spacing and alignment to the formation of MNTs22. Optical tweezers on microfluidic chip were used to pull out MNTs23.…”

The Micropillar Structure on Silk Fibroin Film Influence Intercellular Connection Mediated by Nanotubular Structures

“…Furthermore, 3D analysis revealed that nanotubular structures between cells connected at their nearest distance without contact with substrate and were aligned mostly parallel to the substrate surface [22]. The results confirmed that TNTs can span a long distance by hovering in the culture medium without contact with substrate.…”

“…TNT is a thin dynamic filopodia-like membrane protrusion connecting surrounding cells over long distances. Different from the structure of filopodia, TNTs have been described as stretched membrane channels between interconnected cells at their nearest distance, meanwhile they hover freely in the cell culture medium and do not contact with the substratum on flat surface [3,22]. However, the extension and connection of TNTs on microstructured surface is unclear.…”

“…Interestingly, TNTs hover freely in the cell culture medium and do not contact with the substratum, and they were stretched between interconnected cells attached at their nearest distance [3]. 3D analysis further showed that nanotubular structure between cells at their nearest distance has no contact to the substrate and are aligned mostly parallel to the surface [22]. These results demonstrated that TNTs can span a long distance by hovering extension and do not contact with substrate in the flat culture system.…”