Cellnet technology to generate 3D, functional, single-cell networks in custom architectures within collagen

Area of Science:

• Biotechnology
• Cell Biology
• Biomaterials

Background:

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

Purpose of the Study:

• To introduce Cellnet, a novel technology for generating user-defined 3D single-cell networks in custom architectures.
• To demonstrate the ability to study cell network functionality and signaling responses in real-time.

Main Methods:

• Utilizing natural collagen crosslinked in microfluidic chips.
• Employing femtosecond laser-assisted cavitation to create 3D microchannel networks.
• Seeding cells into microchannels for self-organization into defined 3D networks.

Main Results:

• Successfully generated viable, interconnected 3D single-cell networks in various architectures (grid, circle, lines, spiral).
• Demonstrated the creation of heterotypic Cellnets with multiple cell types.
• Enabled real-time monitoring of calcium signaling and signal propagation in response to stimuli.
• Showcased the ability to generate disrupted Cellnets and analyze signaling dynamics.

Conclusions:

• Cellnet technology offers a reproducible and accurate method for creating organized 3D single-cell networks.
• This platform facilitates the study of cell network behavior and responses to stimuli.
• Cellnet has broad potential applications in fundamental and applied biosciences.