Amyotrophic Lateral Sclerosis (ALS), is a neurodegenerative disorder characterised by the progressive and rapid loss of upper and lower motor neurons within the central nervous system. ALS exhibits significant heterogeneity, demonstrating variability in clinical presentation, disease progression, and the neural populations affected. This diversity has historically complicated identifying disease mechanisms and devising universally effective treatments. Organoids derived from patient pluripotent stem cells offer an innovative platform to investigate ALS, by recapitulating the multicellular complexity and key features of human neuron development and function in vitro. This study generates cortical and spinal motor neuron organoids using pluripotent stem cells obtained from patients with familial C9orf72 and TARDBP mutations. Cortical and spinal organoids demonstrated precise cellular regional specifications and sustained growth and functionality long-term. Preliminary results revealed that neurons within patient-derived organoids displayed key phenotypical hallmarks of ALS pathology, including a reduction in the complexity and density of their dendritic arborization, swollen soma, and increased cellular death. When compared to healthy isogenic controls, patient spinal organoids demonstrated an elevated presence of S100β and GFAP-positive astrocytes, indicating potential regional and disease specific alterations. Our initial findings suggest that physiologically relevant environments are pivotal in unravelling multicellular interactions required for understanding ALS pathogenesis. This insight may contribute towards the development of patient-specific therapies, including high-throughput drug discovery targeted at specific, and potentially subtle, early disease profiles.