Cross-site reproducibility of human cortical organoids reveals consistent cell type composition and architecture Article Swipe
Madison R. Glass
,
Elisa A. Waxman
,
Satoshi Yamashita
,
Michael J. Lafferty
,
A. Hernández Beltrán
,
Tala M. Farah
,
Niyanta K Patel
,
Nana Matoba
,
Sara M. Ahmed
,
Mary Srivastava
,
Emma Drake
,
Liam T. Davis
,
Meghana Yeturi
,
Kexin Sun
,
Michael I. Love
,
Kazue Hashimoto‐Torii
,
Deborah L. French
,
Jason L. Stein
·
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.1101/2023.07.28.550873
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.1101/2023.07.28.550873
Background: Reproducibility of human cortical organoid (hCO) phenotypes remains a concern for modeling neurodevelopmental disorders. While guided hCO protocols reproducibly generate cortical cell types in multiple cell lines at one site, variability across sites using a harmonized protocol has not yet been evaluated. We present an hCO cross-site reproducibility study examining multiple phenotypes. Methods: Three independent research groups generated hCOs from one induced pluripotent stem cell (iPSC) line using a harmonized miniaturized spinning bioreactor protocol. scRNA-seq, 3D fluorescent imaging, phase contrast imaging, qPCR, and flow cytometry were used to characterize the 3 month differentiations across sites. Results: In all sites, hCOs were mostly cortical progenitor and neuronal cell types in reproducible proportions with moderate to high fidelity to the in vivo brain that were consistently organized in cortical wall-like buds. Cross-site differences were detected in hCO size and morphology. Differential gene expression showed differences in metabolism and cellular stress across sites. Although iPSC culture conditions were consistent and iPSCs remained undifferentiated, primed stem cell marker expression prior to differentiation correlated with cell type proportions in hCOs. Conclusions: We identified hCO phenotypes that are reproducible across sites using a harmonized differentiation protocol. Previously described limitations of hCO models were also reproduced including off-target differentiations, necrotic cores, and cellular stress. Improving our understanding of how stem cell states influence early hCO cell types may increase reliability of hCO differentiations. Cross-site reproducibility of hCO cell type proportions and organization lays the foundation for future collaborative prospective meta-analytic studies modeling neurodevelopmental disorders in hCOs.
Related Topics
Concepts
Induced pluripotent stem cell
Cell type
Biology
Progenitor cell
Stem cell
Cell
Organoid
Phenotype
Reproducibility
Cell biology
Neuroscience
Computational biology
Embryonic stem cell
Chemistry
Gene
Genetics
Chromatography
Metadata
- Type
- preprint
- Language
- en
- Landing Page
- https://doi.org/10.1101/2023.07.28.550873
- https://www.biorxiv.org/content/biorxiv/early/2023/07/29/2023.07.28.550873.full.pdf
- OA Status
- green
- Cited By
- 2
- References
- 89
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4385382128
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4385382128Canonical identifier for this work in OpenAlex
- DOI
-
https://doi.org/10.1101/2023.07.28.550873Digital Object Identifier
- Title
-
Cross-site reproducibility of human cortical organoids reveals consistent cell type composition and architectureWork title
- Type
-
preprintOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2023Year of publication
- Publication date
-
2023-07-29Full publication date if available
- Authors
-
Madison R. Glass, Elisa A. Waxman, Satoshi Yamashita, Michael J. Lafferty, A. Hernández Beltrán, Tala M. Farah, Niyanta K Patel, Nana Matoba, Sara M. Ahmed, Mary Srivastava, Emma Drake, Liam T. Davis, Meghana Yeturi, Kexin Sun, Michael I. Love, Kazue Hashimoto‐Torii, Deborah L. French, Jason L. SteinList of authors in order
- Landing page
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https://doi.org/10.1101/2023.07.28.550873Publisher landing page
- PDF URL
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https://www.biorxiv.org/content/biorxiv/early/2023/07/29/2023.07.28.550873.full.pdfDirect link to full text PDF
- Open access
-
YesWhether a free full text is available
- OA status
-
greenOpen access status per OpenAlex
- OA URL
-
https://www.biorxiv.org/content/biorxiv/early/2023/07/29/2023.07.28.550873.full.pdfDirect OA link when available
- Concepts
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Induced pluripotent stem cell, Cell type, Biology, Progenitor cell, Stem cell, Cell, Organoid, Phenotype, Reproducibility, Cell biology, Neuroscience, Computational biology, Embryonic stem cell, Chemistry, Gene, Genetics, ChromatographyTop concepts (fields/topics) attached by OpenAlex
- Cited by
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2Total citation count in OpenAlex
- Citations by year (recent)
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2025: 1, 2024: 1Per-year citation counts (last 5 years)
- References (count)
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89Number of works referenced by this work
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10Other works algorithmically related by OpenAlex
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| publication_date | 2023-07-29 |
| publication_year | 2023 |
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