Early Drug Discovery Unit (EDDU), Montreal Neurological Institute-Hospital, Department of Neurology and Neurosurgery, McGill University, 3801 University Street, Montreal, Quebec H3A 2B4, Canada.
Early Drug Discovery Unit (EDDU), Montreal Neurological Institute-Hospital, Department of Neurology and Neurosurgery, McGill University, 3801 University Street, Montreal, Quebec H3A 2B4, Canada.
Methods. 2022 Jul;203:465-477. doi: 10.1016/j.ymeth.2021.07.008. Epub 2021 Jul 24.
By providing a three-dimensional in vitro culture system with key features of the substantia nigra region in the brain, 3D neuronal organoids derived from human induced pluripotent stem cells (iPSCs) provide living neuronal tissue resembling the midbrain region of the brain. However, a major limitation of conventional brain organoid culture is that it is often labor-intensive, requiring highly specialized personnel for moderate throughput. Additionally, the methods published for long-term cultures require time-consuming maintenance to generate brain organoids in large numbers. With the increasing need for human midbrain organoids (hMOs) to better understand and model Parkinson's disease (PD) in a dish, there is a need to implement new workflows and methods to both generate and maintain hMOs, while minimizing batch to batch variation. In this study, we developed a method with microfabricated disks to scale up the generation of hMOs. This opens up the possibility to generate larger numbers of hMOs, in a manner that minimizes the amount of labor required, while decreasing variability and maintaining the viability of these hMOs over time. Taken together, producing hMOs in this manner opens up the potential for these to be used to further PD studies.
通过提供具有大脑黑质区域关键特征的三维体外培养系统,源自人诱导多能干细胞(iPSC)的 3D 神经元类器官提供了类似于大脑中脑区域的活神经元组织。然而,传统脑类器官培养的一个主要限制是它通常劳动强度大,需要高度专业化的人员才能实现中等通量。此外,已发表的用于长期培养的方法需要耗时的维护才能大量生成脑类器官。随着在培养皿中更好地理解和模拟帕金森病(PD)的需求不断增加,需要实施新的工作流程和方法来生成和维持 hMO,同时最大限度地减少批次间的差异。在这项研究中,我们开发了一种使用微加工盘的方法来扩大 hMO 的生成规模。这为生成更多数量的 hMO 提供了可能性,这种方法最大限度地减少了所需的劳动力,同时降低了变异性并保持了这些 hMO 的活力随时间的推移。总之,以这种方式生成 hMO 为进一步研究 PD 提供了可能性。