3D Polymer Architectures for the Identification of Optimal Dimensions for Cellular Growth of 3D Cellular Models

Abstract

Organ-on-chips and scaffolds for tissue engineering are vital assay tools for pre-clinical testing and prediction of human response to drugs and toxins, while providing an ethical sound replacement for animal testing. A success criterion for these models is the ability to have structural parameters for optimized performance. Here we show that two-photon polymerization fabrication can create 3D test platforms, where scaffold parameters can be directly analyzed by their effects on cell growth and movement. We design and fabricate a 3D grid structure, consisting of wall structures with niches of various dimensions for probing cell attachment and movement, while providing easy access for fluorescence imaging. The 3D structures are fabricated from bio-compatible polymer SZ2080 and subsequently seeded with A549 lung epithelia cells. The seeded structures are imaged with confocal microscopy, where spectral imaging with linear unmixing is used to separate auto-fluorescence scaffold contribution from the cell fluorescence. The volume of cellular material present in different sections of the structures is analyzed, to study the influence of structural parameters on cell distribution. Furthermore, time-lapse studies are performed to map the relation between scaffold parameters and cell movement. In the future, this kind of differentiated 3D growth platform, could be applied for optimized culture growth, cell differentiation, and advanced cell therapies.

Keywords

• 3D cell scaffolds
• two-photon polymerization
• spectral imaging
• linear-unmixing
• time-lapse studies