It is recognized that topographical features such as ridges and grooves can dramatically influence cell phenotype, motivating the development of substrates with precisely biomimetic topography for study of the influence on cultured cells. Intestinal basement membrane topography has been precisely replicated using plasma enhanced chemical vapor deposition (CVD) of poly(2-hydroxyethyl methacrylate) (pHEMA) on native tissue. The ability for CVD pHEMA to coat and retain the complex architecture of the intestinal basement membrane at the micrometer scale was demonstrated using electron microscopy and surface chemical analysis (XPS). The suitability of CVD pHEMA as a cell culture substrate was assessed. Caco-2 cells maintained a high (>85%) viability on CVD pHEMA. Cell attachment and proliferation on CVD pHEMA were similar to those observed on materials traditionally used for cell culture and microfabrication purposes. Results indicate that CVD pHEMA is useful for development of precise (micrometer-scale) topographically biomimetic substrates for cell culture.