understanding the relationship between the cell and the substrate in microenvironments is a critical issue in cell biology research. In this paper, we report the response of HS68 normal human foreskin fibroblast cells to three-dimensional silicon microstructures, which are designed and fabricated using a single-mask fabrication technique. Our device composed of a network of microfluidic channels and microchambers with different widths and depths. Our results show that human fibroblast cells do not tend to go inside microchambers having isotropic cross sections and curved sidewalls, and moreover, their growth rate decreases as the depth increases. The growth rate decline has been utilized as a new method to create patterned cell culture.