人类诱导多能干细胞（iPSC）衍生的大脑类器官有可能重现大脑发育的最早阶段，作为研究正常大脑发育和疾病的有效体外模型。然而，当前的脑类器官培养方法面临着一些挑战，包括通量低、类器官生成的高变异性以及在整个培养过程中细胞在水凝胶中的耗时、多次转移和封装。这些限制阻碍了脑类器官的广泛应用，包括在临床和工业实验室环境中对化合物进行高通量评估。在这项研究中，我们展示了一种在柱板平台上从 iPSC 生成多个脑类器官的直接方法，消除了劳动密集型、多次转移和封装步骤的需要，以确保脑类器官的可重复生成。我们在超低附着 (ULA) 384 孔板中形成胚状体 (EB)，然后使用简单的夹心和倒置方法将它们转移到含有基质胶的柱板中。立柱板上的每个立柱都包含一个球体，球体转移的成功率在 95 - 100% 范围内。通过区分柱板上的 EB，我们实现了变异系数 (CV) 低于 19% 的大脑类器官的稳健生成。值得注意的是，我们的球体转移方法与柱板相结合，可以实现大脑类器官的小型化培养，减轻了类器官变异性的问题，并且与 6-/ 中的传统类器官培养相比，通过允许原位类器官评估，有可能显着提高检测通量。 24 孔板、培养皿和旋转烧瓶。

Human induced pluripotent stem cell (iPSCs)-derived brain organoids have potential to recapitulate the earliest stages of brain development, serving as an effective in vitro model for studying both normal brain development and disorders. However, current brain organoid culture methods face several challenges, including low throughput, high variability in organoid generation, and time-consuming, multiple transfer and encapsulation of cells in hydrogels throughout the culture. These limitations hinder the widespread application of brain organoids including high-throughput assessment of compounds in clinical and industrial lab settings. In this study, we demonstrate a straightforward approach of generating multiple cerebral organoids from iPSCs on a pillar plate platform, eliminating the need for labor-intensive, multiple transfer and encapsulation steps to ensure the reproducible generation of cerebral organoids. We formed embryoid bodies (EBs) in an ultra-low attachment (ULA) 384-well plate and subsequently transferred them to the pillar plate containing Matrigel, using a straightforward sandwiching and inverting method. Each pillar on the pillar plate contains a single spheroid, and the success rate of spheroid transfer was in a range of 95 - 100%. By differentiating the EBs on the pillar plate, we achieved robust generation of cerebral organoids with a coefficient of variation (CV) below 19%. Notably, our spheroid transfer method in combination with the pillar plate allows miniaturized culture of cerebral organoids, alleviates the issue of organoid variability, and has potential to significantly enhance assay throughput by allowing in situ organoid assessment as compared to conventional organoid culture in 6-/24-well plates, petri dishes, and spinner flasks.