Generation of cerebral organoids from human pluripotent stem cells

Nat Protoc. 2014 Oct;9(10):2329-40. doi: 10.1038/nprot.2014.158. Epub 2014 Sep 4.


Human brain development exhibits several unique aspects, such as increased complexity and expansion of neuronal output, that have proven difficult to study in model organisms. As a result, in vitro approaches to model human brain development and disease are an intense area of research. Here we describe a recently established protocol for generating 3D brain tissue, so-called cerebral organoids, which closely mimics the endogenous developmental program. This method can easily be implemented in a standard tissue culture room and can give rise to developing cerebral cortex, ventral telencephalon, choroid plexus and retinal identities, among others, within 1-2 months. This straightforward protocol can be applied to developmental studies, as well as to the study of a variety of human brain diseases. Furthermore, as organoids can be maintained for more than 1 year in long-term culture, they also have the potential to model later events such as neuronal maturation and survival.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Brain / cytology
  • Brain / growth & development*
  • Humans
  • Neuroepithelial Cells / physiology
  • Organoids / growth & development*
  • Pluripotent Stem Cells*
  • Time Factors
  • Tissue Culture Techniques / instrumentation
  • Tissue Culture Techniques / methods*