Nanoporous alumina membranes were utilized as cell culture substrates. Two different anodization processes were applied in order to obtain highly ordered, self-supporting nanoporous alumina membranes. The resulting membranes exhibit uniform nanopores with constant diameters having aspect ratios (pore depth:pore diameter) larger than 1000. Tentative experiments evaluating the cellular interaction of a hepatoma cell line (HepG2) with the nanoporous alumina membranes will be described. Furthermore, cell proliferation as well as the adhesion behavior of the cells on the porous substrates was investigated. Scanning electron microscopy (SEM) and focused ion beam (FIB) technology were applied to examine the cell morphology. The experiments revealed that the nanoporous membranes can be used as cell culture substrates and that they have no adverse effects on cell proliferation and cell viability. Besides, cells on membranes with pore diameters larger than 200 nm developed small cell extensions (filopodia) which penetrated into the nanopores.