The extracellular matrix (ECM) plays important roles in influencing cellular behavior such as attachment, differentiation, and proliferation. However, in conventional culture and tissue engineering strategies, single proteins are frequently utilized, which do not mimic the complex extracellular microenvironment seen in vivo. In this study we report a method to decellularize brain tissue using detergents. This decellularized brain matrix is rich in glycosaminoglycans and contains collagen I, collagen III, collagen IV, collagen V, collagen VI, perlecan, and laminin. By further processing the material into a liquid form, the brain matrix can be used as a cell culture coating. Neurons derived from human induced pluripotent stem cells plated on the brain matrix express neuronal markers and assume neuronal morphology. Additionally, the same material can potentially be used as a scaffold for tissue engineering as it reassembles upon injection in vivo to form a gel. Thus, our work demonstrates the ability to use decellularized brain ECM for cell culture and tissue engineering applications.