We have investigated the culture of human embryonic stem cells (hESCs) on interfaces of the thermotropic liquid crystal, TL205, that are decorated with thin films of the extracellular matrix, Matrigel. hESCs seeded at the liquid-crystal/Matrigel interface survive for weeks, and cell colonies grow over this time. The cells show levels of differentiation comparable to that observed for cells on Matrigel-coated glass controls. Polarized and fluorescence microscopy reveal that the orientational order of the liquid crystal is coupled to the presence and organization of Matrigel. This enables straightforward imaging of the reorganization of Matrigel by the hESCs through changes in the appearance of the liquid crystal when observed using polarized light microscopy. The coupling between Matrigel and TL205 thus provides a simple tool for monitoring the reorganization of the Matrigel film over time. Our results suggest new approaches to the culture of cells and measurements of cell–extracellular-matrix interactions.