Generating LGE/CGE Organoids from Human Pluripotent Stem Cell-derived Brain Organoids Without External Signal Induction

Human pluripotent stem cell (hPSC)-derived GABAergic neurons offer potential for treating neurological disorders by restoring disrupted inhibitory circuits, yet current differentiation methods show limited efficiency, purity, and subtype specificity. We present an approach for generating lateral/caudal ganglionic eminence (LGE/CGE) organoids (LCOs) from hPSC-derived brain organoids (BOs) without external signal induction. LCOs bud from the BO surface and are predominantly composed of LGE/CGE-type GABAergic neurons that mature into functional inhibitory neurons. LCOs and BOs exhibit distinct subtype compositions: LCOs contain both LGE-type neurons, with the capacity to form striatal medium spiny neurons, and abundant CGE-type neurons, whereas BOs contain fewer CGE-type neurons. Single-cell transcriptomic analysis reveals that LCOs closely resemble human embryonic LGE/CGE neurons at gestational weeks 12-13. We also developed a method to selectively enrich CXCR4⁺ CGE-type neurons from LCOs. This platform enables efficient generation of human LGE/CGE-type GABAergic neurons for disease modeling and cell therapy development.