Deriving early single-rosette brain organoids from human pluripotent stem cells

Andrew M Tidball; Wei Niu; Qianyi Ma; Taylor N Takla; J Clayton Walker; Joshua L Margolis; Sandra P Mojica-Perez; Roksolana Sudyk; Lu Deng; Shannon J Moore; Ravi Chopra; Vikram G Shakkottai; Geoffrey G Murphy; Yukun Yuan; Lori L Isom; Jun Z Li; Jack M Parent

doi:10.1016/j.stemcr.2023.10.020

Deriving early single-rosette brain organoids from human pluripotent stem cells

Stem Cell Reports. 2023 Dec 12;18(12):2498-2514. doi: 10.1016/j.stemcr.2023.10.020. Epub 2023 Nov 22.

Authors

Andrew M Tidball¹, Wei Niu¹, Qianyi Ma², Taylor N Takla¹, J Clayton Walker¹, Joshua L Margolis¹, Sandra P Mojica-Perez¹, Roksolana Sudyk¹, Lu Deng¹, Shannon J Moore¹, Ravi Chopra³, Vikram G Shakkottai⁴, Geoffrey G Murphy⁵, Yukun Yuan⁶, Lori L Isom⁷, Jun Z Li⁸, Jack M Parent⁹

Affiliations

¹ Department of Neurology, University of Michigan Medical School, Ann Arbor, MI, USA.
² Department of Human Genetics, University of Michigan Medical School, Ann Arbor, MI, USA.
³ Department of Neurology, Washington University School of Medicine, Saint Louis, MO, USA.
⁴ Department of Neurology, University of Texas Southwestern Medical Center, Dallas, TX, USA.
⁵ Department of Molecular and Integrative Physiology, University of Michigan Medical School, Ann Arbor, MI, USA; Michigan Neuroscience Institute, University of Michigan Medical School, Ann Arbor, MI, USA.
⁶ Department of Pharmacology, University of Michigan Medical School, Ann Arbor, MI, USA.
⁷ Department of Neurology, University of Michigan Medical School, Ann Arbor, MI, USA; Department of Molecular and Integrative Physiology, University of Michigan Medical School, Ann Arbor, MI, USA; Department of Pharmacology, University of Michigan Medical School, Ann Arbor, MI, USA.
⁸ Department of Human Genetics, University of Michigan Medical School, Ann Arbor, MI, USA; Department of Computational Medicine and Bioinformatics, University of Michigan Medical School, Ann Arbor, MI, USA.
⁹ Department of Neurology, University of Michigan Medical School, Ann Arbor, MI, USA; Michigan Neuroscience Institute, University of Michigan Medical School, Ann Arbor, MI, USA; VA Ann Arbor Healthcare System, Ann Arbor, MI, USA. Electronic address: parent@med.umich.edu.

Abstract

Brain organoid methods are complicated by multiple rosette structures and morphological variability. We have developed a human brain organoid technique that generates self-organizing, single-rosette cortical organoids (SOSR-COs) with reproducible size and structure at early timepoints. Rather than patterning a 3-dimensional embryoid body, we initiate brain organoid formation from a 2-dimensional monolayer of human pluripotent stem cells patterned with small molecules into neuroepithelium and differentiated to cells of the developing dorsal cerebral cortex. This approach recapitulates the 2D to 3D developmental transition from neural plate to neural tube. Most monolayer fragments form spheres with a single central lumen. Over time, the SOSR-COs develop appropriate progenitor and cortical laminar cell types as shown by immunocytochemistry and single-cell RNA sequencing. At early time points, this method demonstrates robust structural phenotypes after chemical teratogen exposure or when modeling a genetic neurodevelopmental disorder, and should prove useful for studies of human brain development and disease modeling.

Keywords: PCDH19; cortical spheroid; dorsal forebrain; epilepsy; induced pluripotent stem cells; mosaicism; neural tube defects; neurodevelopment; neurulation; valproic acid.

MeSH terms

Brain
Cell Differentiation
Humans
Induced Pluripotent Stem Cells* / metabolism
Organoids
Pluripotent Stem Cells*

Abstract

MeSH terms

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