This dissertation aims to further the understanding of the complex communication that occurs as cells interact with topographical and chemical patterns on a biomaterial interface. The research accomplishes this through two aims fabricating cell substrate surface topography and chemical patterns independently using non-cleanroom approaches, and analyzing higher order cellular response to surface features. The work will impact biomaterial surface modification and fabrication which will apply to biomedical implanted devices, tissue engineering scaffolds, and biological analysis devices. The first aim seeks to apply non-traditional topographical and chemical patterning methods in order to create independent topographical and chemical patterns on cell culture substrates. Experiments use the resulting patterned substrates to quantify cellular alignment to surface topography and compare the relative influence of topographical and chemical patterns on cellular response. The combined patterning methods of imprint lithography and micro-contact printing result in a high-throughput technique applicable to a variety of materials and a range of feature sizes from nanoscale through microscale, thereby enabling future analysis of cell response to surface features. The second aim evaluates the impact of topographical and chemical features on cellular differentiation. Experiments use patterned topography overlaid with a characterized chemical model layer to evaluate the effects of topography on myoblast differentiation and alignment. Chemical patterns that independently control available cell spreading area and modulate cell-cell contact are used to investigate the impact of cell-cell contact on differentiation.