In this study, the polymer thin films containing micropatterns and biological functionalities were created by one-step procedure. The adjustable compositions among poly(methyl methacrylate) (PMMA), solvents, nonsolvent, and additional macromolecules formed the polymer thin films with different diameters ranging from 5 to 37 ?m. The influences of topographical and chemical cues were investigated by directly cultivating L-929 fibroblasts on the prepared samples. The results revealed the predominant effect of surface topography that the cell density of L-929 fibroblasts increased proportionally with the average diameter of microconcaves. The cell number raised significantly on the PMMA thin films containing type I collagen and dopamine, with or without microstructures.On the other hand, the addition of bovine serum albumin in PMMA limited the growth of cells. The surface chemical composition and cell responses were evaluated by electron spectroscopy for chemical analysis (ESCA), viability assay, and immunostaining, respectively. This work proposed a simple and effective approach to incorporate the biological functions and surface topography simultaneously onto surface of materials that provided further applications for biomedical materials.