Variation of human neural stem cells generating organizer states in vitro before committing to cortical excitatory or inhibitory neuronal fates [human RNAseq #1]

Better understanding the progression of neural stem cells (NSCs) in the developing cerebral cortex is important for modeling neurogenesis and defining the pathogenesis of neuropsychiat-ric disorders. Here we used RNA-sequencing, cell imaging and lineage tracing of mouse and human in vitro NSCs to model the generation of cortical neuronal fates. We show that con-served signaling mechanisms regulate the acute transition from proliferative NSCs to commit-ted glutamatergic excitatory neurons. As human telencephalic NSCs developed from pluripo-tentcy in vitro, they first transitioned through organizer states that spatially pattern the cortex before generating glutamatergic precursor fates. NSCs derived from multiple human pluripotent lines varied in these early states leading differentially to dorsal or ventral telencephalic fates. This work furthers systematic analysis of the earliest patterning events that generate the major neuronal trajectories of the human telencephalon. The gene amplitudes for the GWCoGAPS patterns in this data can be found in the supplemental files. Overall design: Of the 3 series in this super series, this covers the passaging of human H9 ESC-derived neural stem cells (hNSCs) in vitro with varying exposure to FGF2 during final passage (all time points are 6 days after final passage - used in Figures 3 & 4).