Human tissues are sophisticated ensembles of various cell types embedded in the complex but defined structures of the extracellular matrix (ECM). ECM is configured in a hierarchical structure from nano- to microscale, with many biological molecules forming large scale configurations and textures with feature sizes up to macroscopic scale (several hundred microns). The physicochemical, biological and mechanostructural properties of native ECM play a critical role in constructing a microenvironment for cells and tissues. In conjunction with the rapid evolution of material science and its fabrication techniques, studies of the topography and elasticity of ECM and other materials have allowed advanced interrogation of cellular mechanotransduction and cellular responses to mechanostructural cues. By learning from and mimicking the highly organized ECM structures found in vivo, topography-guided approaches to regulate cell function and fate have been widely investigated in the last several decades. Here, we review recent efforts in mimicking the micro- and nanotopography of the native ECM in vitro for the regulation of cellular behaviors. We also discuss how these biomimetic topographical surfaces have been applied to fundamental cell mechanobiology studies into cell adhesions, migrations, and differentiation as well as toward efforts in tissue engineering.