Single-nucleus Transcriptomics Identifies Stem Cell-Derived Graft Composition in a Model of Cortical Stroke

Human cortical neural progenitor cell transplantation holds significant potential in the treatment of cortical stroke by replacing lost cortical neurons and repairing damaged brain circuits. However, commonly utilized human cortical neural progenitors are limited in their ability to yield a substantial proportion of diverse cortical neurons and require an extended period to achieve functional maturation and synaptic integration, potentially diminishing the optimal therapeutic benefits of cell transplantation for cortical stroke. Here, we generated forkhead box G1 (FOXG1)-positive forebrain progenitors from human inducible pluripotent stem cells by optimizing the established differentiation method. Furthermore, we performed an analysis using single-nucleus RNA sequencing (snRNA-seq) combined with comprehensive histological and functional analyses to characterize intracortical grafts from human induced pluripotent stem cells after functional maturation in a preclinical rat stroke model. We show that after transplantation, these FOXG1-positive neural progenitor cells (NPCs) have the ability to differentiate into a high proportion of balanced upper- and deep-layer cortical neurons, as well as inhibitory neurons, exhibiting early functional maturation. These findings provide systematic and compelling evidence for the suitability of these FOXG1 progenitors for neuronal replacement in ischemic cortical stroke.