Generation of Multicue Cellular Microenvironments by UV-Photopatterning of Three-Dimensional Cell Culture Substrates

Abstract

The extracellular matrix is an important regulator of cell function. Environmental cues existing in the cellular microenvironment, such as ligand distribution and tissue geometry, have been increasingly shown to play critical roles in governing cell phenotype and behavior. However, these environmental cues and their effects on cells are often studied separately using in vitro platforms that isolate individual cues, a strategy that heavily oversimplifies the complex in vivo situation of multiple cues.Engineering approaches can be particularly useful to bridge this gap, by developing experimental setups that capture the complexity of the in vivo microenvironment, yet retain the degree of precision and manipulability of in vitro systems.This study highlights an approach combining ultraviolet (UV)-based protein patterning and lithography-based substrate microfabrication, which together enable highthroughput investigation into cell behaviors in multicue environments. By means of maskless UV-photopatterning, it is possible to create complex, adhesive protein distributions on three-dimensional (3D) cell culture substrates on chips that contain a variety of well-defined geometrical cues. The proposed technique can be employed for culture substrates made from different polymeric materials and combined with adhesive patterned areas of a broad range of proteins. With this approach, single cells, as well as monolayers, can be subjected to combinations of geometrical cues and contact guidance cues presented by the patterned substrates. Systematic research using combinations of chip materials, protein patterns, and cell types can thus provide fundamental insights into cellular responses to multicue environments.

Control of hydrostatic pressure and osmotic stress in 3D cell culture for mechanobiological studies

“…Integrating this basis with recent breakthroughs in tissue fabrication, biophysics, and biomaterials, should enable bottom-up strategies for the design of bioreactors that are well-suited to investigate the mechanobiology of HP and OS. Novel methods in 3D tissue bioprinting [159][160][161][162][163], mechanical characterization of biological tissues [164][165][166][167][168][169], the formulation of responsive biomaterials with tunable biomechanical and biochemical properties [170][171][172][173][174][175][176], and the development of 3D organotypic cell cultures [177][178][179][180][181][182] highlight a battery of innovations that can be exploited towards the design of advanced materials systems and bioreactors for the control of HP and OS.…”

Stimuli-Responsive Hydrogels: The Dynamic Smart Biomaterials of Tomorrow

“…Studies showed that cell attachment of primary human keratocytes and human dermal fibroblasts could be influenced by these protein contact-guidance cues but also geometrical cues such as the substrate curvature. The reported photopatterning technique in combination with digital masks enabled the formation of various geometries with a resolution as small as 1.5 μm . Recently, Falandt and colleagues reported the use of volumetric bioprinting to draw and imprint gradients and patterns of growth factors in any custom-designed geometry across a centimeter-scale …”

“…The reported photopatterning technique in combination with digital masks enabled the formation of various geometries with a resolution as small as 1.5 μm. 125 Recently, Falandt and colleagues reported the use of volumetric bioprinting to draw and imprint gradients and patterns of growth factors in any custom-designed geometry across a centimeter-scale. 126 In conclusion, light-triggered methods are very interesting for the local modification of photoresponsive hydrogels as it can be delivered with very high precision and thus good resolution.…”