Human stem cell-derived GABAergic interneuron development reveals early emergence of subtype diversity followed by gradual electrochemical maturation [scRNA-seq]

Medial ganglionic eminence-derived inhibitory GABAergic pallial interneurons (MGE-pINs) are essential for cortical development and function. Dysfunction in MGE-pINs is associated with numerous neurological disorders. We developed a human MGE-pIN cell therapy candidate from pluripotent stem cells, which is undergoing clinical trial evaluation for drug-resistant epilepsy (NCT05135091). Here, we performed single nuclei RNA sequencing and analyzed the transcriptomes of xenografted hMGE-pINs over the lifespan of host mice. Comparative transcriptomic analyses with published human brain datasets revealed that over 97% of grafted human cells developed into pallial MGE-derived somatostatin (SST) and parvalbumin (PVALB) subtypes, including populations that exhibit selective vulnerability in Alzheimer’s disease. Transplanted hMGE-pINs progressed through distinct transcriptional states sequentially involving neuronal migration, synapse organization, and the maturation of electrophysiological properties, demonstrating a surprisingly rapid emergence of subclass-specific features within weeks after grafting. We present molecular, electrophysiological, and morphological data that collectively confirm the derivation of diverse bona-fide human SST and PVALB subtypes, providing a high-fidelity model to study human MGE-pIN development and functional maturation in both healthy and diseased states as well as a compositional atlas for regenerative cell therapy applications. Human ESCs were patterned into MGE projenitors and then differentiated into postmitotic pallial GABAergic interneurons. Cell identity and lot compostion at the end of process (EOP) was analyzed by using 10X Genomics scRNA-seq platform.