It is becoming recognised that traditional methods of culture in vitro on flat substrates do not replicate physiological conditions well, and a number of studies have indicated that the physical environment is crucial to the directed functioning of cells in vivo. In this paper we report the development of a platform with cell-like features that is suitable for in vitro investigation of cell activity. Biological cells were imprinted in hard methacrylate copolymer using soft lithography. The cell structures were replicated at high nanometre scale resolution, as confirmed by atomic force microscopy. Optimisation of the methacrylate-based copolymer mixture for transparency and biocompatibility was performed, and cytotoxicity and chemical stability of the cured polymer in cell culture conditions were evaluated. Cells of an endometrial adenocarcinoma cell line (Ishikawa) were cultured on bioimprinted substrates. The cells exhibited differential attachment on the bioimprint substrate surface compared to those on areas of flat surface and preferentially followed the pattern of the original cell footprint. The results revealed for the first time that the cancer cells distinguished between behavioural cues from surfaces that had features reminiscent of themselves and that of flat areas. Therefore the imprinted platform will lend itself to detailed studies of relevant physical substrate environments on cell behaviour. The material is not degraded and its permanency allows reuse of the same substrate in multiple experimental runs. It is simple and does not require expensive or specialised equipment. In this work cancer cells were studied, and the growth behaviour of the tumour-derived cells was modified by alterations of the cells’ physical environment. Implications are also clear for studies in other crucial areas of health, such as wound healing and artificial tissues.