Here we present long-term patterning and response studies of neuroblasts on arrays of extracellular matrix (ECM) protein lines fabricated by cracking a polymer-supported thin film and selectively depositing proteins inside the cracks. Neuroblasts cultured for five days on the protein matrix switched between spreading and retraction upon changing the protein line width. This technique provides a novel tool to pursue further understanding of the basic mechanisms of neuroblast pathfinding on adhesion and morphology. Biomedical device design will also benefit from materials engineered with the reconfigurable protein patterns.