Table_9_Brainstem Organoids From Human Pluripotent Stem Cells.xlsx

The brainstem is a posterior region of the brain, composed of three parts, midbrain, pons, and medulla oblongata. It is critical in controlling heartbeat, blood pressure, and respiration, all of which are life-sustaining functions, and therefore, damages to or disorders of the brainstem can be lethal. Brain organoids derived from human pluripotent stem cells (hPSCs) 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 are limited in the extent hPSCs recapitulate human brain development and hence are not able to fully elucidate the diseases affecting various components of the brain such as brainstem. Here, we developed a method to generate human brainstem organoids (hBSOs), containing midbrain/hindbrain progenitors, noradrenergic and cholinergic neurons, dopaminergic neurons, and neural crest lineage cells. Single-cell RNA sequence (scRNA-seq) analysis, together with evidence from proteomics and electrophysiology, revealed that the cellular population in these organoids was similar to that of the human brainstem, which raises the possibility of making use of hBSOs in investigating central nervous system disorders affecting brainstem and in efficient drug screenings.

脑干（brainstem）是大脑的后部区域，由中脑（midbrain）、脑桥（pons）和延髓（medulla oblongata）三部分组成，在调控心跳、血压与呼吸等维持生命的核心生理功能中至关重要，因此脑干损伤或功能紊乱可危及生命。源自人类多能干细胞（human pluripotent stem cells, hPSCs）的大脑类器官（brain organoids）能够重现人类大脑发育过程，有望应用于中枢神经系统（central nervous system）疾病的医学研究。然而，现有类器官模型在人类多能干细胞重现大脑发育的程度上存在局限，因此无法全面阐明累及脑干等大脑多种结构的疾病机制。本研究开发了一种制备人类脑干类器官（human brainstem organoids, hBSOs）的方法，此类类器官包含中脑/后脑祖细胞（midbrain/hindbrain progenitors）、去甲肾上腺素能神经元（noradrenergic neurons）、胆碱能神经元（cholinergic neurons）、多巴胺能神经元（dopaminergic neurons）以及神经嵴谱系细胞（neural crest lineage cells）。单细胞RNA测序（single-cell RNA sequence, scRNA-seq）分析结合蛋白质组学（proteomics）与电生理学（electrophysiology）证据表明，此类类器官中的细胞群与人类脑干的细胞群高度相似，这为利用hBSOs研究累及脑干的中枢神经系统疾病以及开展高效药物筛选提供了可能。