The extensive use of iron oxide nanomaterials in biomedical applications has prompted the development of a novel substrate for evaluating cell behaviour. This study examines the fabrication of tuneable length iron oxide pillar arrays using the porous nanochannels of anodic aluminium oxide membranes, and evaluates the biocompatibility of the substrate. The electroformed iron pillars were found to conform to the template channels with slightly larger iron oxide pillar diameters, due to the presence of an oxide shell. The biocompatibility was then confirmed with WST-1 proliferation and viability assay of cultured KT98 murine neural/progenitor stem cells on the surface of the pillar array; with no significant difference observed between viable cells after seven days of culture on iron oxide pillars, flat iron oxide, and tissue culture polystyrene. The physical properties of the pillar arrays were linked to the adhesion and spreading of the cells, and found that cells cultured on the pillar arrays had reduced spreading in comparison to tissue culture polystyrene control. In addition, it was found that protein expression was unaffected by culture on iron oxide substrates. The results of this study indicate that iron oxide pillar arrays are suitable to extended cell studies.