TGFβ superfamily signaling regulates the state of human stem cell pluripotency and capacity to create well-structured telencephalic organoids

Stem Cell Reports. 2022 Oct 11;17(10):2220-2238. doi: 10.1016/j.stemcr.2022.08.013. Epub 2022 Sep 29.


Telencephalic organoids generated from human pluripotent stem cells (hPSCs) are a promising system for studying the distinct features of the developing human brain and the underlying causes of many neurological disorders. While organoid technology is steadily advancing, many challenges remain, including potential batch-to-batch and cell-line-to-cell-line variability, and structural inconsistency. Here, we demonstrate that a major contributor to cortical organoid quality is the way hPSCs are maintained prior to differentiation. Optimal results were achieved using particular fibroblast-feeder-supported hPSCs rather than feeder-independent cells, differences that were reflected in their transcriptomic states at the outset. Feeder-supported hPSCs displayed activation of diverse transforming growth factor β (TGFβ) superfamily signaling pathways and increased expression of genes connected to naive pluripotency. We further identified combinations of TGFβ-related growth factors that are necessary and together sufficient to impart broad telencephalic organoid competency to feeder-free hPSCs and enhance the formation of well-structured brain tissues suitable for disease modeling.

Keywords: brain organoid; cerebral cortex; choroid plexus; differentiation; embryonic stem cell; ganglionic eminence; hippocampus; neural development; neural stem cell; neurogenesis; pluripotency; pluripotent stem cell; stem cell heterogeneity.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Differentiation / physiology
  • Humans
  • Organoids* / metabolism
  • Pluripotent Stem Cells* / metabolism
  • Telencephalon / metabolism
  • Transforming Growth Factor beta / metabolism


  • Transforming Growth Factor beta