pp 1-12 | Cite as

A Simple Method of Generating 3D Brain Organoids Using Standard Laboratory Equipment

Part of the Methods in Molecular Biology book series


3D brain organoids are a powerful tool with prospective application for the study of neural development and disease. Here we describe the growth factor-free method of generating cerebral organoids from feeder-dependent or feeder-free human pluripotent stem cells using standard laboratory equipment. The protocol outlined below allows generation of 3D tissues, which replicate human early in vivo brain development up to the end of the first trimester, both in terms of morphology and gene expression pattern.


Brain In vitro Neural differentiation Neurobiology Neurodevelopment Organoids Stem cells Three-dimensional 


  1. 1.
    Eiraku M, Watanabe K, Matsuo-Takasaki M et al (2008) Self-organized formation of polarized cortical tissues from ESCs and its active manipulation by extrinsic signals. Cell Stem Cell 3:519–532CrossRefPubMedGoogle Scholar
  2. 2.
    Paşca AM, Sloan SA, Clarke LE et al (2015) Functional cortical neurons and astrocytes from human pluripotent stem cells in 3D culture. Nat Methods 12:671–678CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Kadoshima T, Sakaguchi H, Nakano T et al (2013) Self-organization of axial polarity, inside-out layer pattern, and species-specific progenitor dynamics in human ES cell-derived neocortex. Proc Natl Acad Sci U S A 110:20284–20289CrossRefPubMedPubMedCentralADSGoogle Scholar
  4. 4.
    Lancaster MA, Renner M, Martin C et al (2013) Cerebral organoids model human brain development and microcephaly. Nature 501:373–379CrossRefPubMedADSGoogle Scholar
  5. 5.
    Kelava I, Lancaster MA (2016) Dishing out mini-brains: current progress and future prospects in brain organoid research. Dev Biol 420(2):199–209CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Lancaster MA, Knoblich JA (2014) Generation of cerebral organoids from human pluripotent stem cells. Nat Protoc 9:2329–2340CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Camp JG, Badsha F, Florio M et al (2015) Human cerebral organoids recapitulate gene expression programs of fetal neocortex development. Proc Natl Acad Sci U S A 112:15672–15677CrossRefPubMedPubMedCentralADSGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.MRC Laboratory of Molecular BiologyCambridgeUK

Personalised recommendations