Midbrain-like Organoids from Human Pluripotent Stem Cells Contain Functional Dopaminergic and Neuromelanin-Producing Neurons

• Published in: Cell stem cell Vol. 19; no. 2; pp. 248 - 257
• Main Authors: Jo, Junghyun, Xiao, Yixin, Sun, Alfred Xuyang, Cukuroglu, Engin, Tran, Hoang-Dai, Göke, Jonathan, Tan, Zi Ying, Saw, Tzuen Yih, Tan, Cheng-Peow, Lokman, Hidayat, Lee, Younghwan, Kim, Donghoon, Ko, Han Seok, Kim, Seong-Oh, Park, Jae Hyeon, Cho, Nam-Joon, Hyde, Thomas M., Kleinman, Joel E., Shin, Joo Heon, Weinberger, Daniel R., Tan, Eng King, Je, Hyunsoo Shawn, Ng, Huck-Hui
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 04.08.2016
• Subjects: Cell Differentiation; Cell Line; Dopaminergic Neurons - metabolism; Humans; Melanins - metabolism; Mesencephalon - cytology; Organoids - cytology; Pluripotent Stem Cells - cytology; Transcription, Genetic
• ISSN: 1934-5909; 1875-9777
• DOI: 10.1016/j.stem.2016.07.005

Recent advances in 3D culture systems have led to the generation of brain organoids that resemble different human brain regions; however, a 3D organoid model of the midbrain containing functional midbrain dopaminergic (mDA) neurons has not been reported. We developed a method to differentiate human pluripotent stem cells into a large multicellular organoid-like structure that contains distinct layers of neuronal cells expressing characteristic markers of human midbrain. Importantly, we detected electrically active and functionally mature mDA neurons and dopamine production in our 3D midbrain-like organoids (MLOs). In contrast to human mDA neurons generated using 2D methods or MLOs generated from mouse embryonic stem cells, our human MLOs produced neuromelanin-like granules that were structurally similar to those isolated from human substantia nigra tissues. Thus our MLOs bearing features of the human midbrain may provide a tractable in vitro system to study the human midbrain and its related diseases. [Display omitted] •Self-organizing midbrain-like organoids (hMLOs) develop from hPSCs in 3D culture•hMLOs, but not mouse MLOs or human cerebral organoids, produce neuromelanin•hMLOs secrete dopamine (DA) and neurons within the hMLOs form functional synapses•Neurons within hMLOs exhibit SNpc DA neuron-like electrophysiological properties Jo et al. report a method for generating human midbrain-like organoids (hMLOs) from hPSCs in 3D culture. The hMLOs contain distinct layers of neuronal cells expressing human midbrain markers, such as neuromelanin, are electrically active, form functional synapses, and produce dopamine, suggesting that they may be useful for studying human midbrain.