CELLNET technology: Spatially organized, functional 3D networks at single cell resolution.

Area of Science:

• Biotechnology
• Cell Biology
• Bioengineering

Background:

• Understanding cellular networks is crucial for fundamental biology and therapeutic development.
• Existing methods struggle to create organized 3D single-cell networks within native extracellular matrix (ECM).

Purpose of the Study:

• To introduce CELLNET, a novel technology for generating interconnected and disrupted 3D single-cell networks in custom configurations within native ECM.
• To demonstrate the capability of CELLNET in creating organized cellular architectures and studying their functional responses.

Main Methods:

• CELLNET involves creating crosslinked collagen within microfluidic devices, followed by femtosecond laser ablation to form 3D microchannel networks.
• Cells are seeded into these networks, allowing migration, self-organization, and formation of interconnected 3D structures.
• The technology allows for the creation of heterotypic networks and user-defined network disruptions.

Main Results:

• CELLNET successfully generated viable, interconnected 3D cell networks in various custom architectures (e.g., grid, spiral).
• Real-time calcium signaling and signal propagation within these networks were monitored under different stimuli.
• Disrupted networks were created, enabling analysis of signaling dynamics following targeted cell injury.

Conclusions:

• CELLNET offers a reproducible and versatile platform for creating organized 3D single-cell networks within native ECM.
• This technology facilitates the study of cell network functionality and response to stimuli, overcoming limitations of current methods.
• CELLNET has broad potential applications in fundamental and applied biosciences, independent of cell type or ECM composition.