CELLNET technology: spatially organized, functional 3D single-cell networks using laser templated collagen

Cells possess the remarkable ability to generate tissue-specific 3D interconnected networks and respond to a wide range of stimuli. Understanding the link between the spatial arrangement of individual cells and their networks’ emergent properties is necessary for the discovery of both fundamental biology as well as applied therapeutics. However, current methods are unable to generate interconnected and organized single cell 3D networks within native extracellular matrix (ECM). To address this challenge, we report a novel technology coined as CELLNET. This involves the generation of crosslinked collagen within multi-chambered microfluidic devices followed by femtosecond laser ablation of 3D microchannel networks and cell seeding. Using model cells, we show that cell migrate within ablated networks within hours, self-organize and form viable, interconnected, 3D networks in custom architectures. Heterotypic CELLNETs can also be generated. Real-time calcium signaling response in normal and laser ablated or disrupted CELLNETs can also be investigated. CELLNET is agnostic of cell types, ECM formulations, 3D cell-connectivity designs, could pave the way to address a range of applications.