Brain organoid methods are complicated by multiple rosette structures and morphological variability. We have developed a human brain organoid technique that generates self-organizing, single-rosette cortical organoids (SOSR-COs) with reproducible size and structure at early timepoints. Rather than patterning a 3-dimensional embryoid body, we initiate brain organoid formation from a 2-dimensional monolayer of human pluripotent stem cells patterned with small molecules into neuroepithelium and differentiated to cells of the developing dorsal cerebral cortex. This approach recapitulates the 2D to 3D developmental transition from neural plate to neural tube. Most monolayer fragments form spheres with a single central lumen. Over time, the SOSR-COs develop appropriate progenitor and cortical laminar cell types as shown by immunocytochemistry and single-cell RNA sequencing. At early time points, this method demonstrates robust structural phenotypes after chemical teratogen exposure or when modeling a genetic neurodevelopmental disorder, and should prove useful for studies of human brain development and disease modeling.

脑类器官方法因多种玫瑰花结结构和形态变异而变得复杂。我们开发了一种人脑类器官技术，可生成自组织、单莲座皮质类器官（SOSR-CO），在早期时间点具有可重复的大小和结构。我们不是构建 3 维胚状体的图案，而是从用小分子图案化的二维单层人类多能干细胞开始脑类器官的形成，形成神经上皮，并分化为发育中的背侧大脑皮层的细胞。这种方法概括了从神经板到神经管的 2D 到 3D 发育转变。大多数单层碎片形成具有单个中心腔的球体。随着时间的推移，SOSR-CO 会发育出适当的祖细胞和皮质层状细胞类型，如免疫细胞化学和单细胞 RNA 测序所示。在早期时间点，该方法在化学致畸剂暴露后或在模拟遗传性神经发育障碍时表现出强大的结构表型，并且应该对人类大脑发育和疾病建模的研究有用。