Brainstem organoids from human pluripotent stem cells contain neural crest population

Brainstem controls heartbeat, blood pressure and respiration which are vital for life, therefore, damage to brainstem can be lethal. Brain organoids derived from human pluripotent stem cells recapitulate the course of human brain development and are expected to be useful for medical research on central nervous system disorders. However, existing organoid models have limitations, hampering the elucidation of diseases affecting specific components of the brain. Here, we developed a method to generate human brainstem organoids (hBSOs), containing neural crest stem cells as well as midbrain/hindbrain progenitors, noradrenergic and cholinergic neurons, and dopaminergic neurons, demonstrated by specific electrophysiological signatures. Single-cell RNA-sequence analysis, together with proteomics and electrophysiology further revealed that the cellular population in the organoids is similar to that of the human brainstem and neural crest, which makes it possible to make use of the hBSOs for modeling and grafting for transplantation. Overall design: Single-cell RNA-sequencing of human brainstem organoids