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Individual brain organoids reproducibly form cell diversity of the human cerebral cortex
Open accessAbstract
Experimental models of the human brain are needed for basic understanding of its development and disease 1. Human brain organoids hold unprecedented promise for this purpose; however, they are plagued by high organoid-to-organoid variability 2,3. This has raised doubts as to whether developmental processes of the human brain can occur outside the context of embryogenesis with a degree of reproducibility that is comparable to the endogenous tissue. Here we show that an organoid model of the dorsal forebrain can reliably generate a rich diversity of cell types appropriate for the human cerebral cortex. We performed single-cell RNA-sequencing analysis of 166,242 cells isolated from 21 individual organoids, finding that 95% of the organoids generate a virtually indistinguishable compendium of cell types, following similar developmental trajectories and with a degree of organoid-to-organoid variability comparable to that of individual endogenous brains. Furthermore, organoids derived from different stem cell lines show consistent reproducibility in the cell types produced. The data demonstrate that reproducible development of the complex cellular diversity of the central nervous system does not require the context of the embryo, and that establishment of terminal cell identity is a highly constrained process that can emerge from diverse stem cell origins and growth environments.
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“Furthermore, several studies have shown that guided cortical organoid differentiation yields highly reproducible organoids as compared to unguided organoid differentiation (72–74).”
From Understanding monocyte-driven neuroinflammation in Alzheimer’s disease using human cortical organoid microphysiological systems (2025) · cites this paper as [74]
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“Organoid development followed the expected temporal changes (Lancaster et al., 2013; Velasco et al., 2019; Watanabe et al., 2017; Yoon et al., 2019; Figures 1B–1D; Figure S1A).”
From ELAVL4, splicing, and glutamatergic dysfunction precede neuron loss in MAPT mutation cerebral organoids (2021) · cites this paper as Velasco et al., 2019
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“scRNA-seq analysis of human intestinal and brain organoids used random forest classifiers to identify the cell types in their organoid cultures;73,74”
From Single-cell transcriptomics of human-skin-equivalent organoids (2023) · cites this paper as [74]
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“All cell sub-types were present in WA25 and DSP-GFP organoids, but in different proportions (Fig. 2b, c), suggesting further protocol optimization may be needed to reduce variability19.”
From Hair-bearing human skin generated entirely from pluripotent stem cells (2020) · cites this paper as [19]
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Metadata
- Authors
- Velasco, S., Kedaigle, A. J., Simmons, S. K., Nash, A., Rocha, M., Quadrato, G., Paulsen, B., Nguyen, L., Adiconis, X., Regev, A., Levin, J. Z., & Arlotta, P.
- Affiliations
- Broad Institute (US)
- Corresponding author
- Arlotta, Paola
- ORCID
- Venue
- Nature
- Published
- 2019-06
- Volume
- 570
- Issue
- 7762
- Pages
- 523-527
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC6906116
- OpenAlex
- W2949020598
- Cited by
- 1,094
- References
- 39
- Citations by year
- 2026: 145 · 2025: 156 · 2024: 184 · 2023: 131 · 2022: 148 · 2021: 142 · 2020: 150 · 2019: 35
- FWCI
- 44.71
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Digital Access to Scholarship at Harvard (DASH) (Harvard University) (acceptedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- Funders
- Howard Hughes Medical Institute
- Broad Institute
- Stanley Center for Psychiatric Research, Broad Institute
- Klarman Cell Observatory, Broad Institute
- National Institute of Mental Health (P50MH094271, U01MH115727, R01MH112940)
- Topics
- Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
- Keywords
- human brain organoids, cortical development, cell type diversity, dorsal forebrain, stem cell origin, reproducibility, single-cell transcriptomics
- MeSH terms
- RNA-Seq, Cell Line, Cerebral Cortex, Female, Fetus, Humans, Male, Organoids, Time Factors, Reproducibility of Results, Prosencephalon, Tissue Culture Techniques, Induced Pluripotent Stem Cells, Single-Cell Analysis, Transcriptome
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