GSE128357
Self-organizing human trunk 3D neuromuscular organoids
Neuromuscular networks assemble during early human embryonic development and are essential for the control of body movement. Previous neuromuscular junction modeling efforts using human pluripotent stem cells (hPSCs) generated either spinal cord neurons or skeletal muscles in monolayer culture. Here, we use hPSC-derived axial stem cells, the building blocks of the posterior body, to simultaneously generate spinal cord neurons and skeletal muscle cells that self-organize to generate human neuromuscular organoids (NMOs) that can be maintained in 3D for several months. Single-cell RNA-sequencing of individual organoids revealed reproducibility across experiments and enabled the tracking of the neural and mesodermal differentiation trajectories as organoids developed and matured. NMOs contain functional neuromuscular junctions supported by terminal Schwann cells. They contract and develop central pattern generator-like neuronal circuits. Finally, we successfully use NMOs to recapitulate key aspects of myasthenia gravis pathology, thus highlighting the significant potential of NMOs for modeling neuromuscular diseases in the future. Overall design: We have analyzed single cells from 10 organoids at Day 5. The organoids were separated in two samples (5 pooled organoids each) and labelled with different indexes. In total 5,135 cells were analyzed with a median of 3,333 genes per cell. For Day 50 we analyzed 4 organoids. One sample consisted of one organoid analyzed independently, and the other Day 50 sample consisted of 3 organoids tagged with different indexes. In total 17,294 cells were analyzed with a median of 2,343 genes per cell.