This work introduces a novel process for the fabrication of polymer cell culture substrates containing physical topography based on timepoint specific cell phenotype replicas. Bioimprinting of human nasal chondrocyte at different cell adhesion time points was used to demonstrate the nanoscale replication process. Atomic force microscopy confirmed morphology progression at 1, 6, 12, and 24 h timepoints corresponding to dedifferentiation of the chondrocytes to fibroblast-like phenotype. Topographical analysis of the imprinted substrates yielded an inverse relationship between surface coverage, increasing from 11 to 87, and maximum average pattern depth, decreasing from 1.2 μm to 430 nm. Methacrylate bioimprint features were successively replicated into a transitional polydmethylsiloxane mold used as an intermediate for further replication into polystyrene by a high-throughput embossing method. Substrates fabricated with this process have applications in stem cell engineering, regenerative medicine, and implantable devices. © 2012 American Vacuum Society.