A Defined Xeno-Free Matrix Supports Midbrain Dopaminergic Cell Differentiation

Stem cell-based therapy holds great promise for treating Parkinson's disease (PD), yet its clinical translation depends on safely and efficiently generating midbrain dopaminergic cells (mDACs) in vitro. One major challenge in current protocols is the safety risks associated with extracellular matrix (ECM) components. Commonly used materials such as laminin may contain undefined animal-derived components, while their human recombinant alternatives face technical and economic limitations, highlighting an urgent need for safe and effective materials for scalable clinical-grade cell manufacturing. Sialic acid is recognized as a crucial nutrient during neural development and a key structural component of brain sialylglycopeptide and gangliosides, but its use to optimize in vitro neuronal differentiation protocols has not been assessed. In this study, starting from the observation that sialyllactose supports mDAC differentiation, we identified a series of sialyl-glycopeptides as candidate matrix materials and comparatively evaluated their supramolecular morphology, nanomechanical flexibility, and matrix-supporting performance. Among these candidates, SL-4F, replacing laminin, combined with poly-l-ornithine and fibronectin, promotes robust mDAC growth and differentiation. The efficacy and safety of this formulation are confirmed in vitro and in vivo, suggesting SL-4F as a synthetic and safer alternative for clinical-grade cell culture for PD and, potentially, other stem cell-based therapies.