3D cellular self-assembly on optical disc-imprinted nanopatterns†

Three-dimensional (3D) cellular assemblies, such as cancer spheroids and organoids, are increasingly valued for their physiological relevance, and versatility in biological applications. Nanopatterns that mimic the extracellular matrix provide crucial topological cues, creating a physiologically relevant cellular environment and guiding cellular behaviors. However, the high cost and complex, time-consuming nature of the nanofabrication process have limited the widespread adoption of nanopatterns in diverse biological applications. In this study, we present a straightforward and cost-effective elastomer replica molding method utilizing commercially available optical discs to generate various nanopatterns, such as nanogroove/ridge, nanoposts, and nanopits, varying in spacing and heights. Using the nanopatterned well chips (NW-Chips), we demonstrated the efficient formation of 3D multicellular self-assemblies of three different types of cancer cells. Our findings highlight the accessibility and affordability of optical discs as tools for nanopattern generation, offering promising avenues for modulating cell behaviors and advancing diverse biological applications.