The development of observing the effects of culture environment on cell bioactivity is important for deeply understanding fundamental physiological issues in various in vitro experiments. Meanwhile, as we know, surfaces with multiscale structures on cell culture substrates could be used to aid in induction of many cellular responses, which are related to the advancement of tissue engineering and regenerative medicine. Herein, we review morphological and functional responses of different types of cells cultured in a two-dimensional or three-dimensional culture substrates.