Generation of Brain Region-specific Organoids using a Miniaturized Spinning Bioreactor and Modelling ZIKV Exposure. Homo sapiens

Cerebral organoids, three-dimensional cultures that model organogenesis, provide a new platform to investigate human brain development. High cost, variability and tissue heterogeneity limit accessibility and broad applications of current organoid technologies. Here we developed a miniaturized spinning bioreactor (SpinΩ) to generate forebrain-specific organoids from human iPSCs. These organoids recapitulate key features of human cortical development, including progenitor zone organization, neurogenesis, gene expression, and importantly, a distinct human-specific outer radial glia cell layer. We have also developed protocols to generate midbrain and hypothalamic organoids. Finally, we employed this forebrain organoid platform to model Zika virus (ZIKV) exposure. Quantitative analyses revealed that preferential, productive ZIKA infection of cortical neural progenitors leads to increased cell death and reduced proliferation, resulting in decreased neuronal cell layer volume that resembles microcephaly. Together, our brain region-specific organoids and SpinΩ provide an accessible and versatile platform for modeling human brain development and diseases, and for compound testing. Overall design: Time course of human cerebral organoid cultures. No Zika virus infection is involved.

大脑类器官（cerebral organoids）是模拟器官发生过程的三维培养体系，为研究人类大脑发育提供了全新的技术平台。现有类器官技术受限于高昂的成本、批次间差异以及组织异质性，其可及性与应用范围均受到限制。本研究开发了一款微型旋转生物反应器（SpinΩ），可利用人类诱导多能干细胞（induced pluripotent stem cells, iPSCs）制备前脑特异性类器官。此类类器官可重现人类大脑皮层发育的关键特征，包括祖细胞区组织结构、神经发生过程、基因表达模式，尤为重要的是，还能形成人类特有的外放射状胶质细胞层。本研究同时建立了制备中脑与下丘脑类器官的实验方案。最后，本研究利用该前脑类器官平台构建了寨卡病毒（Zika virus, ZIKV）暴露模型。定量分析结果显示，寨卡病毒对皮层神经祖细胞的偏好性增殖感染会引发细胞死亡增多与增殖能力下降，最终导致神经元层体积缩减，呈现出小头畸形的病理特征。综上，本研究开发的脑区特异性类器官体系与SpinΩ生物反应器，为人类大脑发育与疾病建模以及化合物筛选提供了一个易用且多功能的研究平台。实验整体设计：人类大脑类器官的时序培养实验，未涉及寨卡病毒感染操作。