Sivitilli Adam, Ghiasi Parisa, Attisano Liliana
Department of Biochemistry, Donnelly Centre, University of Toronto, Toronto, ON, Canada.
Bio Protoc. 2021 Apr 20;11(8):e3985. doi: 10.21769/BioProtoc.3985.
Recent advances in stem cell technology have allowed researchers to generate 3D cerebral organoids (COs) from human pluripotent stem cells (hPSCs). Indeed, COs have provided an unprecedented opportunity to model the developing human brain in a 3D context, and in turn, are suitable for addressing complex neurological questions by leveraging advancements in genetic engineering, high resolution microscopy, and tissue transcriptomics. However, the use of this model is limited by substantial variations in the overall morphology and cellular composition of organoids derived from the same pluripotent cell line. To address these limitations, we established a robust, high-efficiency protocol for the production of consistent COs by optimizing the initial phase of embryoid body (EB) formation and neural induction. Using this protocol, COs can be reproducibly generated with a uniform size, shape, and cellular composition across multiple batches. Furthermore, organoids that developed over extended periods of time (3-6 months) showed the establishment of relatively mature features, including electrophysiologically active neurons, and the emergence of oligodendrocyte progenitors. Thus, this platform provides a robust experimental model that can be used to study human brain development and associated disorders. Graphic abstract: Overview of cerebral organoid development from pluripotent stem cells.
干细胞技术的最新进展使研究人员能够从人多能干细胞（hPSC）中生成3D脑类器官（CO）。事实上，CO为在3D环境中模拟发育中的人类大脑提供了前所未有的机会，进而适用于通过利用基因工程、高分辨率显微镜和组织转录组学的进展来解决复杂的神经学问题。然而，该模型的使用受到来自同一多能细胞系的类器官在整体形态和细胞组成上存在显著差异的限制。为了解决这些限制，我们通过优化胚状体（EB）形成和神经诱导的初始阶段，建立了一种稳健、高效的方案来生产一致的CO。使用该方案，可以在多个批次中以均匀的大小、形状和细胞组成可重复地生成CO。此外，长时间（3 - 6个月）发育的类器官显示出相对成熟特征的建立，包括具有电生理活性的神经元，以及少突胶质细胞祖细胞的出现。因此，该平台提供了一个强大的实验模型，可用于研究人类大脑发育及相关疾病。图形摘要：多能干细胞来源的脑类器官发育概述
