Abstract
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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
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 3:519–532
Paşca AM, Sloan SA, Clarke LE et al (2015) Functional cortical neurons and astrocytes from human pluripotent stem cells in 3D culture. Nat Methods 12:671–678
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 110:20284–20289
Lancaster MA, Renner M, Martin C et al (2013) Cerebral organoids model human brain development and microcephaly. Nature 501:373–379
Kelava I, Lancaster MA (2016) Dishing out mini-brains: current progress and future prospects in brain organoid research. Dev Biol 420(2):199–209
Lancaster MA, Knoblich JA (2014) Generation of cerebral organoids from human pluripotent stem cells. Nat Protoc 9:2329–2340
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 112:15672–15677
Acknowledgments
We would like to thank all members of the Lancaster Lab for helpful discussion. Research in the Lancaster Lab is supported by the MRC (MC_UP_1201/9) as well as a 3Rs prize from the NC3Rs.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
Sutcliffe, M., Lancaster, M.A. (2017). A Simple Method of Generating 3D Brain Organoids Using Standard Laboratory Equipment. In: Turksen, K. (eds) Organoids. Methods in Molecular Biology, vol 1576. Humana, New York, NY. https://doi.org/10.1007/7651_2017_2
Download citation
DOI: https://doi.org/10.1007/7651_2017_2
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-7616-4
Online ISBN: 978-1-4939-7617-1
eBook Packages: Springer Protocols