Encapsulation of Cells in a Collagen Matrix Surrounded by an Alginate Hydrogel Shell for 3D Cell Culture

Area of Science:

• Biotechnology
• Biomaterials Engineering
• Cell Biology

Background:

• Mammalian cell culture techniques aim to replicate in vivo 3D tissue structures.
• Protein-based matrices allow for self-organized multicellular assemblies.
• High-throughput screening and standardized methods require controlled cell encapsulation and monitoring.

Purpose of the Study:

• To design and develop a novel microfluidic method for creating 3D cell culture compartments.
• To create submillimeter core-shell structures with alginate hydrogel shells and collagen-based cell-laden cores.
• To enable controlled cell encapsulation and 3D tissue formation compatible with pipetting tools.

Main Methods:

• Utilized a microfluidic device for high-speed co-extrusion in air to form a compound jet.
• Controlled jet fragmentation to produce core-shell liquid drops.
• Collected drops in a gelling bath to rapidly harden the alginate shell and allow for collagen self-assembly.

Main Results:

• Successfully formulated the core solution to maintain cell viability under physiological conditions.
• Prevented flow disturbances detrimental to the jetting method.
• Encapsulated Caco-2 cells proliferated and formed a polarized epithelial monolayer with apical membrane facing the medium.

Conclusions:

• The developed microfluidic method enables the creation of 3D cell culture compartments with controlled cell encapsulation.
• The core-shell structures support cell proliferation and the formation of functional polarized epithelial monolayers.
• This technique offers a promising approach for high-throughput screening and standardized 3D cell culture.