Understanding the mechanism of how surface topography influences mammalian cells is important for the development of medical implants and tissue engineering. Although variety of cell types have been used in cell-substrate studies for different purposes, hepatocytes have been paid relatively less attention. In this study, we investigated the influences of nanopores and microgrooves on the morphology and migration of hepatoma cell line, BEL-7402 cells. The cells were cultured on different synthetic substrates including flat cell culture plate, flat silicon wafer sputtered with alumina, nanoporous (140 nm in diameter) anodized aluminum membrane (AAM) surface, flat polydimethylsiloxane (PDMS), and PDMS patterned with microgrooves of various widths (10 μm, 30 μm and 50 μm in width, and 2 μm in depth). Cellular behaviour on these surfaces was studied using fluorescent microscopy, time lapse microscopy and scanning electron microscopy (SEM)