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bioRxiv · 10.1101/2023.02.10.528087

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

Zhou, Z.·Kakegawa, W.·Fujimori, K.·Sho, M.·Shimamura, R.·Supakul, S.·Yoshimatsu, S.·Kohyama, J.·Yuzaki, M.·Okano, H.
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Abstract

Cortical excitatory neurons (Cx neurons) are the most dominant neuronal cell type in the cerebral cortex, which 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 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 efficient induction of Cx neurons from the feeder-free-conditioned human embryonic stem cells (ES cells) and induced pluripotent stem cells (iPS cells). By using this method, human ES/iPS cells could be differentiated into ~99% MAP2-positive neurons by three weeks, and these induced neurons, within five weeks, presented various characteristics of mature excitatory neurons such as strong expression of glutamatergic neuron-specific markers (subunits of AMPA and NDMA receptors and CAMKII), highly synchronized spontaneous firing and excitatory postsynaptic current (EPSC). Moreover, the Cx neurons showed susceptibility to the toxicity of A{beta}42 oligomers and excitotoxicity of excessive glutamates, which is another advantage in terms of toxicity test and searching for the therapeutic agents. Taken together, this study provides a novel research platform for the study of neural development and degeneration based on the feeder-free human ES/iPS cell system.

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Zhou, Z., Kakegawa, W., Fujimori, K., Sho, M., Shimamura, R., Supakul, S., Yoshimatsu, S., Kohyama, J., Yuzaki, M., Okano, H.. 2023-02-12. Highly efficient induction of functionally mature excitatory neurons from feeder-free human ES/iPS cells. https://doi.org/10.1101/2023.02.10.528087

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