Generation of striatal neurons from human induced pluripotent stem cells by controlling extrinsic signals with small molecules

• Published in: Stem cell research 2021-08, Vol.55, p.102486-102486, Article 102486
• Main Authors: Amimoto, Naoya, Nishimura, Kaneyasu, Shimohama, Shun, Takata, Kazuyuki
• Format: Article
• Language: English
• Subjects: Human induced pluripotent stem cells; Lateral ganglionic eminence; Medium spiny neurons; Regional patterning; Striatal neurons
• ISSN: 1873-5061; 1876-7753
• DOI: 10.1016/j.scr.2021.102486

•Large-scale preparation of hiPSC-derived LGE progenitors was achieved.•Striatal neurons were generated from LGE progenitors in vitro using small molecules.•Mature hiPSC-derived striatal neurons exhibited spontaneous spiking activity.•Striatal neurospheres were generated that resembled the developing human striatum. Human induced pluripotent stem cells (hiPSCs) are powerful tools for modeling human brain development and treating neurodegenerative diseases. Here we established a robust protocol with high scalability for generating striatal medium spiny neurons (MSNs) from hiPSCs using small molecules under two- and three-dimensional culture conditions. Using this protocol, GSH2+ lateral ganglionic eminence (LGE) progenitors were generated in two-dimensional culture by Sonic hedgehog signaling activation using purmorphamine, WNT signaling inhibition using XAV939, and dual-SMAD inhibition using LDN193189 and A83-01. Quantitative PCR analysis revealed sequential expression of LGE and striatal genes during differentiation. These LGE progenitors subsequently gave rise to DARPP32+ MSNs exhibiting spontaneous and evoked monophasic spiking activity. Applying this protocol in three-dimensional culture, we generated striatal neurospheres with gene expression profiles and cell layer organization resembling that of the developing striatum, including distinct ventricular and subventricular zones and DARPP32+ neurons at the surface. This protocol provides a useful experimental model for studying striatal development and yields cells potentially applicable for regenerative medicine to treat striatum-related disorders such as Huntington’s disease.