Light-Induced Differentiation of Forebrain Organoids by NVOC-SAG.

Hedgehog signaling pathway shapes our body by regulating proliferation and differentiation of cells. The spatial and temporal distribution pattern of its ligands finely controls the activity of the Hedgehog pathway during development. To mimic the active regulation of Hedgehog pathway, we have developed a light-inducible Hedgehog signaling activator 6-nitroveratryloxy-carbonyl Smoothened agonist (NVOC-SAG). Here we describe a method to selectively induce ventral differentiation of human iPS cell-derived forebrain organoids in a light-dependent manner. This article describes preparation of NVOC-SAG, culture of iPS cell-derived forebrain organoids, light irradiation, and downstream analyses.