This article describes a solvent treatment process for transforming standard flat polystyrene surfaces into textured microenvironments for three-dimensional (3D) cell culture. The process can be used with off-the-shelf microplates having 6, 12, 96, or even 384-well formats and completed in a few steps. The surface microtexture resembles tightly packed, non-interconnected micropores having a median pore size of either 115 or 19 μm depending on the strength of the solvent mixture. Hansen solubility parameter analysis was used to map out the polymer–solvent interaction conditions necessary for creating the surface texture. Primary human hepatocytes cultured on two-dimensional control surfaces attached with a flat monolayer morphology while those cultured on surfaces having 115 μm-sized pores exhibited a round 3D morphology. The cells also showed more in-vivo like behavior on the porous surface by exhibiting significantly higher basal enzymatic activity and expressing higher levels of several drug metabolism-related genes. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014, 131, 40181.
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