Improved Protocol for Reproducible Human Cortical Organoids Reveals Early Alterations in Metabolism with MAPT Mutations

• Published in: bioRxiv : the preprint server for biology
• Main Authors: Bertucci, Taylor, Bowles, Kathryn R, Lotz, Steven, Qi, Le, Stevens, Katherine, Goderie, Susan K, Borden, Susan, Oja, Laura Maria, Lane, Keith, Lotz, Ryan, Lotz, Hailey, Chowdhury, Rebecca, Joy, Shona, Arduini, Brigitte L, Butler, David C, Miller, Michael, Baron, Heide, Sandhof, Carl Alexander, Silva, M Catarina, Haggarty, Stephen J, Karch, Celeste M, Geschwind, Daniel H, Goate, Alison M, Temple, Sally
• Format: Journal Article
• Language: English
• Published: United States 11.07.2023

Cerebral cortical-enriched organoids derived from human pluripotent stem cells (hPSCs) are valuable models for studying neurodevelopment, disease mechanisms, and therapeutic development. However, recognized limitations include the high variability of organoids across hPSC donor lines and experimental replicates. We report a 96-slitwell method for efficient, scalable, reproducible cortical organoid production. When hPSCs were cultured with controlled-release FGF2 and an SB431542 concentration appropriate for their / expression level, organoid cortical patterning and reproducibility were significantly improved. Well-patterned organoids included 16 neuronal and glial subtypes by single cell RNA sequencing (scRNA-seq), frequent neural progenitor rosettes and robust BCL11B+ and TBR1+ deep layer cortical neurons at 2 months by immunohistochemistry. In contrast, poorly-patterned organoids contain mesendoderm-related cells, identifiable by negative QC markers including . Using this improved protocol, we demonstrate increased sensitivity to study the impact of different mutations from patients with frontotemporal dementia (FTD), revealing early changes in key metabolic pathways.