Robust production of uniform human cerebral organoids from pluripotent stem cells

Life Sci Alliance. 2020 Apr 17;3(5):e202000707. doi: 10.26508/lsa.202000707. Print 2020 May.


Human cerebral organoid (hCO) models offer the opportunity to understand fundamental processes underlying human-specific cortical development and pathophysiology in an experimentally tractable system. Although diverse methods to generate brain organoids have been developed, a major challenge has been the production of organoids with reproducible cell type heterogeneity and macroscopic morphology. Here, we have directly addressed this problem by establishing a robust production pipeline to generate morphologically consistent hCOs and achieve a success rate of >80%. These hCOs include both a radial glial stem cell compartment and electrophysiologically competent mature neurons. Moreover, we show using immunofluorescence microscopy and single-cell profiling that individual organoids display reproducible cell type compositions that are conserved upon extended culture. We expect that application of this method will provide new insights into brain development and disease processes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Brain / cytology
  • Brain / metabolism
  • Cell Culture Techniques / methods*
  • Cell Differentiation / physiology
  • Female
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Male
  • Neural Stem Cells / cytology
  • Neurogenesis / physiology
  • Organoids / cytology
  • Organoids / growth & development*
  • Pluripotent Stem Cells / cytology
  • Pluripotent Stem Cells / metabolism*