Cross-site reproducibility of human cortical organoids reveals consistent cell type composition and architecture
While guided human cortical organoid (hCO) protocols reproducibly generate cortical cell types at one site, variability in hCO phenotypes across sites using a harmonized protocol has not yet been evaluated. To determine the cross-site reproducibility of hCO differentiation, three independent research groups assayed hCOs in multiple differentiation replicates from one induced pluripotent stem cell (iPSC) line using a harmonized miniaturized spinning bioreactor protocol across 3 months. hCOs were mostly cortical progenitor and neuronal cell types in reproducible proportions that were consistently organized in cortical wall-like buds. Cross-site differences were detected in hCO size and expression of metabolism and cellular stress genes. Variability in hCO phenotypes correlated with stem cell gene expression prior to differentiation and technical factors associated with seeding, suggesting iPSC quality and treatment are important for differentiation outcomes. Cross-site reproducibility of hCO cell type proportions and organization encourages future prospective meta-analytic studies modeling neurodevelopmental disorders in hCOs.
Authors
- Michael J. Lafferty (ORCID: https://orcid.org/0000-0002-1000-0480)
- Deborah L. French (ORCID: https://orcid.org/0000-0002-7535-1716)
- Jason L. Stein (ORCID: https://orcid.org/0000-0003-4829-0513)
- Alvaro A. Beltran (ORCID: https://orcid.org/0000-0002-1421-3889)
- Tala M. Farah
- Elisa A. Waxman (ORCID: https://orcid.org/0000-0003-4562-7969)
- Michael I. Love (ORCID: https://orcid.org/0000-0001-8401-0545)
- Madison R. Glass
- Satoshi Yamashita (ORCID: https://orcid.org/0000-0002-8609-5797)
- Kexin Sun (ORCID: https://orcid.org/0000-0002-9727-9436)
- Kazue Hashimoto‐Torii (ORCID: https://orcid.org/0000-0003-4001-8259)
- Niyanta K Patel
- Nana Matoba (ORCID: https://orcid.org/0000-0001-5329-0134)
- Emma Drake
- Meghana Yeturi
- Sara M. Ahmed (ORCID: https://orcid.org/0000-0001-8572-3168)
- Mary Srivastava (ORCID: https://orcid.org/0009-0005-2966-6660)
- Liam T. Davis
- Rubal Singla
Institutions
- University of North Carolina at Chapel Hill (US)
- George Washington University Hospital (US)
- University of North Carolina Health Care (US)
- Children's Hospital of Philadelphia (US)
- Children's National (US)
- Jefferson Hospital for Neuroscience (US)
Publication Details
- Journal
- Stem Cell Reports
- Published
- 2024-08-23
- DOI
- https://doi.org/10.1016/j.stemcr.2024.07.008
- Citations
- 19
- Primary Topic
- Pluripotent Stem Cells Research
- Type
- article
- Field-Weighted Citation Impact
- 4.44
Funders
- Foundation of Hope for Research and Treatment of Mental Illness
- National Institutes of Health
- Intellectual and Developmental Disabilities Research Center
- National Institute of Neurological Disorders and Stroke
- Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill
- Eunice Kennedy Shriver National Institute of Child Health and Human Development
- School of Medicine, University of North Carolina at Chapel Hill