Brain organoids offer unprecedented insights into brain development and disease modeling and hold promise for drug screening. Significant hindrances, however, are morphological and cellular heterogeneity, inter-organoid size differences, cellular stress, and poor reproducibility. Here, we describe a method that reproducibly generates thousands of organoids across multiple hiPSC lines. These High Quantity brain organoids (Hi-Q brain organoids) exhibit reproducible cytoarchitecture, cell diversity, and functionality, are free from ectopically active cellular stress pathways, and allow cryopreservation and re-culturing. Patient-derived Hi-Q brain organoids recapitulate distinct forms of developmental defects: primary microcephaly due to a mutation in CDK5RAP2 and progeria-associated defects of Cockayne syndrome. Hi-Q brain organoids displayed a reproducible invasion pattern for a given patient-derived glioma cell line. This enabled a medium-throughput drug screen to identify Selumetinib and Fulvestrant, as inhibitors of glioma invasion in vivo. Thus, the Hi-Q approach can easily be adapted to reliably harness brain organoids’ application for personalized neurogenetic disease modeling and drug discovery.

脑类器官为大脑发育和疾病建模提供了前所未有的见解，并为药物筛选带来了希望。然而，重大障碍是形态和细胞异质性、类器官间大小差异、细胞应激和可重复性差。在这里，我们描述了一种在多个 hiPSC 系中可重复生成数千个类器官的方法。这些高数量脑类器官（Hi-Q 脑类器官）表现出可重复的细胞结构、细胞多样性和功能，不受异位活性细胞应激途径的影响，并允许冷冻保存和再培养。患者来源的 Hi-Q 脑类器官概括了不同形式的发育缺陷：由于 CDK5RAP2 突变引起的原发性小头畸形和 Cockayne 综合征的早衰相关缺陷。Hi-Q 脑类器官对给定的患者来源的神经胶质瘤细胞系显示出可重复的侵袭模式。这使得中等通量药物筛选能够确定 Selumetinib 和 Fulvestrant 作为体内神经胶质瘤侵袭的抑制剂。因此，Hi-Q 方法可以很容易地适应可靠地利用脑类器官的应用进行个性化的神经遗传疾病建模和药物发现。