08-01-2021-11:52 - Brainstem Organoids From Human Pluripotent Stem Cells

Front. Neurosci., 26 June 2020 | https://doi.org/10.3389/fnins.2020.00538

Brainstem Organoids From Human Pluripotent Stem Cells

Nobuyuki Eura1†, Takeshi K. Matsui1,2†, Joachim Luginbühl3†, Masaya Matsubayashi2, Hitoki Nanaura1,2, Tomo Shiota1, Kaoru Kinugawa1, Naohiko Iguchi1, Takao Kiriyama1, Canbin Zheng4, Tsukasa Kouno3, Yan Jun Lan3, Pornparn Kongpracha5, Pattama Wiriyasermkul5, Yoshihiko M. Sakaguchi2, Riko Nagata2, Tomoya Komeda2, Naritaka Morikawa2, Fumika Kitayoshi2, Miyong Jong2, Shinko Kobashigawa2, Mari Nakanishi2, Masatoshi Hasegawa6, Yasuhiko Saito7, Takashi Shiromizu8, Yuhei Nishimura8, Takahiko Kasai9, Maiko Takeda9, Hiroshi Kobayashi10, Yusuke Inagaki11, Yasuhito Tanaka11, Manabu Makinodan12, Toshifumi Kishimoto12, Hiroki Kuniyasu13, Shushi Nagamori5, Alysson R. Muotri14,15, Jay W. Shin3*, Kazuma Sugie1* and Eiichiro Mori2*

• 1Department of Neurology, Nara Medical University, Kashihara, Japan
• 2Department of Future Basic Medicine, Nara Medical University, Kashihara, Japan
• 3Laboratory for Advanced Genomics Circuit, RIKEN Center for Integrative Medical Sciences, Yokohama, Japan
• 4Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX, United States
• 5Laboratory of Biomolecular Dynamics, Department of Collaborative Research, Nara Medical University, Kashihara, Japan
• 6Department of Radiation Oncology, Nara Medical University, Kashihara, Japan
• 7Department of Neurophysiology, Nara Medical University, Kashihara, Japan
• 8Department of Integrative Pharmacology, Graduate School of Medicine, Mie University, Tsu, Japan
• 9Department of Laboratory Medicine and Pathology, National Hospital Organization Kinki-Chuo Chest Medical Center, Sakai, Japan
• 10Department of Obstetrics and Gynecology, Nara Medical University, Kashihara, Japan
• 11Department of Orthopaedic Surgery, Nara Medical University, Kashihara, Japan
• 12Department of Psychiatry, Nara Medical University, Kashihara, Japan
• 13Department of Molecular Pathology, Nara Medical University, Kashihara, Japan
• 14Department of Pediatrics, University of California, San Diego, San Diego, CA, United States
• 15Department of Cellular and Molecular Medicine, University of California, San Diego, San Diego, CA, United States

The brainstem is a posterior region of the brain, composed of three parts, midbrain, pons, and medulla oblongata. It is critical in controlling heartbeat, blood pressure, and respiration, all of which are life-sustaining functions, and therefore, damages to or disorders of the brainstem can be lethal. Brain organoids derived from human pluripotent stem cells (hPSCs) recapitulate the course of human brain development and are expected to be useful for medical research on central nervous system disorders. However, existing organoid models are limited in the extent hPSCs recapitulate human brain development and hence are not able to fully elucidate the diseases affecting various components of the brain such as brainstem. Here, we developed a method to generate human brainstem organoids (hBSOs), containing midbrain/hindbrain progenitors, noradrenergic and cholinergic neurons, dopaminergic neurons, and neural crest lineage cells. Single-cell RNA sequence (scRNA-seq) analysis, together with evidence from proteomics and electrophysiology, revealed that the cellular population in these organoids was similar to that of the human brainstem, which raises the possibility of making use of hBSOs in investigating central nervous system disorders affecting brainstem and in efficient drug screenings.