INFLUENCE OF THE EXTRACELLULAR MICROENVIRONMENT ON NEURONAL DIFFERENTIATION OF URINE-DERIVED STEM CELLS

Urine-derived stem cells (UDSC) are a non-invasive and readily accessible cell source that has gained interest as an in vitro model for neurobiological research. However, the role of the extracellular microenvironment, particularly substrate and matrix composition, in neuronal differentiation remains poorly defined.
In this study, we analysed the neuronal differentiation of UDSC under defined culture conditions, focusing on the contribution of different substrates and extracellular matrices. Neuronal identity was assessed through morphological analysis, immunocytochemistry, and gene expression of neuronal markers.
Our results demonstrate that UDSC can undergo neuronal differentiation, acquiring morphological and molecular neuronal features. Differentiation efficiency, cell morphology, and neuronal marker expression varied significantly depending on the substrate and matrix used, highlighting the strong influence of extracellular cues on cell fate decisions.
Overall, these findings support UDSC as a robust model for studying neuronal differentiation and identify the extracellular microenvironment as a key determinant of neuronal identity, providing a basis for optimizing biomaterial-based in vitro neuronal models.
This work was co-funded by the European Regional Development Fund (ERDF) through the Interreg VI-A Spain–Portugal (POCTEP) 2021–2027 Programme (0145_CROSS_3DTOOL_4ALS); through the Centro 2030 and Centro 2020 Regional Operational Programme under project CENTRO2030-FEDER-00670400 and by the EU Recovery and Resilience Facility and Portuguese national funds via Fundação para a Ciência e a Tecnologia (FCT) under projects LA/P/0058/2020 [DOI: 10.54499/LA/P/0058/2020], UID/PRR/4539/2025 [DOI: 10.54499/UID/PRR/04539/2025], UID/04539/2025, and 2022.00011.CEECIND; additionally, strategic plans and strategic research programmes of excellence from the Regional Government of Castilla y León, co-funded by the ERDF Operational Programme (CLU-2023-1-01).