Highly efficient induction of functionally mature excitatory neurons from feeder-free human ES/iPS cells

• Published in: bioRxiv
• Main Authors: Zhou, Zhi, Kakegawa, Wataru, Fujimori, Koki, Misato Sho, Shimamura, Rieko, Supakul, Sopak, Yoshimatsu, Sho, Kohyama, Jun, Yuzaki, Michisuke, Okano, Hideyuki
• Format: Paper
• Language: English
• Published: Cold Spring Harbor Cold Spring Harbor Laboratory Press 20.02.2023
• Subjects: Alzheimer's disease; Cell differentiation; Cerebral cortex; Cognition; Drug development; Embryo cells; Emotional behavior; Excitatory postsynaptic potentials; Excitotoxicity; Glutamatergic transmission; Neurodegeneration; Neurodegenerative diseases; Neurodevelopmental disorders; Neurons; Pluripotency; Regenerative medicine; Stem cells; Toxicity testing; α-Amino-3-hydroxy-5-methyl-4-isoxazole propionic acid
• DOI: 10.1101/2023.02.10.528087

Cortical excitatory neurons (Cx neurons) are the most dominant neuronal cell type in the cerebral cortex and play a central role in cognition, perception, intellectual behavior, and emotional processing. Robust in vitro induction of Cx neurons may facilitate as a tool for the elucidation of brain development and the pathomechanism of the intractable neurodevelopmental and neurodegenerative disorders, including Alzheimers disease, and thus potentially contribute to drug development. Here, we report a defined method for the efficient induction of Cx neurons from the feeder-free-conditioned human embryonic stem cells (ES cells) and induced pluripotent stem cells (iPS cells). Using this method, human ES/iPS cells could be differentiated into ~99% MAP2-positive neurons by three weeks, and these induced neurons displayed several characteristics of mature excitatory neurons within 5 weeks, such as strong expression of glutamatergic neuron-specific markers (subunits of AMPA and NDMA receptors and CAMKIIα), highly synchronized spontaneous firing and excitatory postsynaptic current (EPSC). In addition, the Cx neurons showed susceptibility to Aβ oligomer toxicity and excessive glutamate excitotoxicity, which is another advantage for toxicity testing and searching for therapeutic agent discovery. Taken together, this study provides a novel research platform for studying neural development and degeneration based on the feeder-free human ES/iPS cell system.Competing Interest StatementH.O. has been a paid scientific advisory board member of San Bio Co., Ltd., and Regenerative Medicine iPS Gateway Center Co. Ltd., and K Pharma, Inc. However, there was no effect of these companies on the interpretation, writing, or publication of this study. H.O. and S.Y. declare no non-financial conflicts of interest with this work. In addition, the other authors declare neither financial nor non-financial conflicts of interest.Footnotes* This version is to retouch the authors' name, correspondence, and orcid.