Generation of Various Telencephalic Regions from Human Embryonic Stem Cells in Three-Dimensional Culture

  • Taisuke Kadoshima
  • Hideya Sakaguchi
  • Mototsugu Eiraku
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1597)

Abstract

In the developing embryo, telencephalon arises from the rostral portion of the neural tube. The telencephalon further subdivides into distinct brain regions along the dorsal-ventral (DV) axis by exogenous patterning signals. Here, we describe a protocol for in vitro generation of various telencephalic regions from human embryonic stem cells (ESCs). Dissociated human ESCs are reaggregated in a low-cell-adhesion 96-well plate and cultured as floating aggregates. Telencephalic neural progenitors are efficiently generated when ESC aggregates are cultured in serum-free medium containing TGFβ inhibitor and Wnt inhibitor. In long-term culture, the telencephalic neural progenitors acquire cortical identities and self-organize a stratified cortical structure as seen in human fetal cortex. By treatment with Shh signal, the telencephalic progenitors acquire ventral (subpallial) identities and generate lateral ganglionic eminence (LGE) and medial ganglionic eminence (MGE). In contrast, by treatment with Wnt and BMP signals, their regional identities shift to more dorsal side that generates choroid plexus and medial palllium (hippocampal primordium).

Key words

SFEBq culture Human ESCs Telencephalon Cerebral cortex Ganglionic eminence Medial pallium Hippocampus 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Taisuke Kadoshima
    • 1
    • 2
  • Hideya Sakaguchi
    • 3
    • 4
  • Mototsugu Eiraku
    • 3
  1. 1.Cell Asymmetry teamRIKEN Center for Developmental BiologyKobeJapan
  2. 2.Asubio Pharma Co., Ltd.KobeJapan
  3. 3.In Vitro Histogenesis teamRIKEN Center for Developmental BiologyKobeJapan
  4. 4.Department of Clinical ApplicationCenter for iPS Cell Research and Application (CiRA), Kyoto UniversityKyotoJapan

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