Cellular adhesiveness to biomaterial is one of the important properties for the success of tissue engineering. The cell–biomaterial interactions involve close cooperation of adhesion proteins, plasma membrane, and cytoskeletons to form focal adhesions during the process of anchoring. Dynamic development of the plasma membrane in the process reflects the cellular biocompatibility and motility. The process of cell attachment beginning from seeding, contact, attachment, and spreading has not been investigated. In this study, we monitored the whole process of cells attaching to the substrate surface by time-lapse confocal microscopy. We observed that the surface configuration of the substratum effects plasma membrane expansion and genomic materials distribution. In contrast to the cells grown on the plate, the cells attached on pillars are with rounded nuclei and with prominent lamellipodia spreading out. Membrane expansion is involved in dynamic development of the plasma membrane and lamellipodia formation for attachment, migration, or proliferation and reflects the cellular physiology status of the cells. This study provides a platform for investigation of cell behavior and dynamic development of subcellular structures regarding cell–biomaterial interactions. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 1149–1154, 2014.