Arrays of transparent rolled-up microtubes can easily be mass-produced using a combination of conventional photolithography, electron beam depositioning, and chemical etching techniques. Here, we culture primary mouse motor neurons and immortalised CAD cells, a cell line derived from the central nervous system, on various microtube substrates to investigate the influence of topographical surface ... [Show full abstract] features on the growth and differentiation behaviour of these cells. Our results indicate that the microtube chips not only support growth of both cell types but also provide a well-defined, geometrically confined 3D cell culture scaffold. Strikingly, our micropatterns act as a platform for axon guidance with protruding cell extensions aligning in the direction of the microtubes and forming complex square-shaped grid-like neurite networks. Our experiments open up a cost-efficient and bio-compatible way of analysing single cell behaviour in the context of advanced micro-/nanostructures with various biological applications ranging from neurite protection studies to cell sensor development.