UV lithography,silicon etching and soft lithography were adopted to fabricate micropillar arrayed cell culture substrates. With this approach,poly(dimethylsiloxane) ( PDMS) topographic substrates with pillar featuring dimensions equal to or larger than 4 μm could be successfully fabricated, with a pillar height of 4 μm. The fabrication of substrates with a pillar feafuring dimension of 2 μm might represent the maximal fabrication capability of this method,as pillar lodging or missing could be found. With a simple tilting angle method,we fabricated a topographic substrate formed with a packed polystyrene microbead array. The arrayed beads could be as small as 2 μm in diameter or even in submicrometer range. Cell culture experiments showed that arrayed topographical substrates can promote adhesion or spreading, affecting resting membrane potential establishement and enhance the responsiveness of voltage-gated calcium channels of neuronal cells. The arrayed topographic substrates provide an efficient tool for engineering the microenvironments for sensor neuronal cells.