Protocol for extraction and characterization of mouse brain-derived extracellular matrix for neuronal cell culture

Summary Neuronal cell cultures are highly sensitive to their microenvironment, particularly the choice of coating substrate. Here, we present a protocol to isolate extracellular matrix (ECM) from mouse brain tissue, providing a native coating material that closely mimics in vivo conditions. We detail decellularization steps, guidelines for measuring ECM quality, and instructions for co-culturing neuronal cells on the resulting substrate. By promoting improved neuronal survival, growth, and differentiation, this protocol has broad implications for in vitro neurobiological research and downstream applications.