A facile method to generate cerebral organoids from human pluripotent stem cells
Keywords:cerebral organoids, human pluripotent stem cell, hanging drop, neural induction protocol, single nuclei RNA-sequencing analysis
Human cerebral organoids (COs) are self-organizing three-dimensional (3D) neural structures that provide a human-specific platform to study the cellular and molecular processes that underlie different neurological events. The first step of CO generation from human pluripotent stem cells (hPSCs) is neural induction, which is an in vitro simulation of neural ectoderm development. Several signaling pathways cooperate during neural ectoderm development and in vitro differentiation of hPSCs toward neural cell lineages is also affected by them. In this study, we considered some of the known sources of these variable signaling cues arising from cell culture media components and sought to modulate their effects by applying a comprehensive combination of small molecules and growth factors for CO generation. Histological analysis demonstrated that these COs recapitulate the neural progenitor zone and early cortical layer organization, containing different types of neuronal and glial cells which was in accordance with single-nucleus transcriptome profiling results. Moreover, patch clamp and intracellular Ca2+ dynamic studies demonstrated that the COs behave as a functional neural network. Thus, this method serves as a facile protocol for generating hPSC-derived COs that faithfully mimic the features of their in vivo counterparts in the developing human brain.
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Copyright (c) 2023 Susan Simorgh, Seyed Ahmad Mousavi, San Kit To, Vincent Pasque, Keimpe Wierda, Tim Vervliet, Meghdad Yeganeh, Paria Pooyan, Yoke Chin Chai, Catherine Verfaillie, Hossein Baharvand
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