Generation of spinal cord organoids from human induced pluripotent stem cells caudalised to a lumbar fate

Organoids offer a powerful platform to model human development and disease in vitro while preserving key features of in vivo tissue architecture and complexity. In this study, we developed a novel, reproducible protocol to generate human induced pluripotent stem cell (iPSC)-derived spinal cord organoids specifically patterned to the lumbar region, a segment rarely modelled in vitro. These organoids recapitulate the ventral spinal cord's cytoarchitecture and exhibit functional neuronal activity, providing a physiologically relevant system for investigating human spinal cord development and neurodegenerative disease mechanisms. Furthermore, we applied 10x Genomics single-cell RNA sequencing to generate the first high-resolution transcriptional map of human lumbar spinal cord organoids, revealing their cellular diversity and molecular signatures. Overall design: Human iPSC-derived spinal cord organoids at day 60 of differentiation were enzymatically dissociated into a single-cell suspension and processed for transcriptomic profiling using the 10x Genomics single-cell RNA sequencing (scRNA-seq) platform.