A facile method to generate cerebral organoids from human pluripotent stem cells (original) (raw)

Authors

Susan Simorgh Department of Developmental Biology, School of Basic Sciences and Advanced Technologies in Biology, University of Science and Culture, Tehran, Iran; Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran; Stem Cell Institute, Department of Development and Regeneration, KU Leuven, Leuven 3000, Belgium https://orcid.org/0000-0002-9347-4728
Seyed Ahmad Mousavi Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran https://orcid.org/0000-0003-3845-3286
San Kit To Department of Development and Regeneration, Lab for Epigenetic Reprogramming, Leuven Stem Cell Institute, Leuven Single-Cell Omics Institute and Leuven Cancer Institute, KU Leuven-University of Leuven, Leuven 3000, Belgium https://orcid.org/0000-0002-3057-2902
Vincent Pasque Department of Development and Regeneration, Lab for Epigenetic Reprogramming, Leuven Stem Cell Institute, Leuven Single-Cell Omics Institute and Leuven Cancer Institute, KU Leuven-University of Leuven, Leuven 3000, Belgium https://orcid.org/0000-0002-5129-0146
Keimpe Wierda VIB-KU Leuven Center for Brain & Disease Research, Leuven 3000, Belgium; Electrophysiology Unit, Leuven 3000, Belgium https://orcid.org/0000-0002-8784-9490
Tim Vervliet Laboratory of Molecular and Cellular Signaling, Department of Cellular and Molecular Medicine, KU Leuven, Leuven 3000, Belgium https://orcid.org/0000-0002-4030-5875
Meghdad Yeganeh Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran https://orcid.org/0009-0005-4738-7761
Paria Pooyan Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran https://orcid.org/0000-0002-1945-1480
Yoke Chin Chai Stem Cell Institute, Department of Development and Regeneration, KU Leuven, Leuven 3000, Belgium; E-mail: yokechin.chai@kuleuven.be https://orcid.org/0000-0001-9913-257X
Catherine Verfaillie Stem Cell Institute, Department of Development and Regeneration, KU Leuven, Leuven 3000, Belgium; E-mail: catherine.verfaillie@kuleuven.be https://orcid.org/0000-0001-7564-4079
Hossein Baharvand Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran; E-mail: h.baharvand@royan-rc.ac.ir https://orcid.org/0000-0001-6528-3687

DOI:

https://doi.org/10.17179/excli2023-6299

Keywords:

cerebral organoids, human pluripotent stem cell, hanging drop, neural induction protocol, single nuclei RNA-sequencing analysis

Abstract

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.

How to Cite

Simorgh, S., Mousavi, S. A., To, S. K., Pasque, V., Wierda, K., Vervliet, T., … Baharvand, H. (2023). A facile method to generate cerebral organoids from human pluripotent stem cells. EXCLI Journal, 22, 1055–1076. https://doi.org/10.17179/excli2023-6299

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