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

• Main Authors: Naoya Amimoto, Kaneyasu Nishimura, Shun Shimohama, Kazuyuki Takata
• Format: Article
• Language: English
• Published: Elsevier 2021-08-01
• Series: Stem Cell Research
• Subjects: Human induced pluripotent stem cells; Medium spiny neurons; Striatal neurons; Lateral ganglionic eminence; Regional patterning
• Online Access: http://www.sciencedirect.com/science/article/pii/S1873506121003330

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.