Purification and characterization of human neural stem and progenitor cells

Abstract

The human brain undergoes rapid development at mid-gestation from a pool of neural stem and progenitor cells (NSPCs) that give rise to the neurons, oligodendrocytes, and astrocytes of the mature brain. Functional study of these cell types has been hampered by a lack of precise purification methods. We describe a method for prospectively isolating ten distinct NSPC types from the developing human brain using cell-surface markers. CD24^-THY1^-/lo cells were enriched for radial glia, which robustly engrafted and differentiated into all three neural lineages in the mouse brain. THY1^hi cells marked unipotent oligodendrocyte precursors committed to an oligodendroglial fate, and CD24⁺THY1^-/lo cells marked committed excitatory and inhibitory neuronal lineages. Notably, we identify and functionally characterize a transcriptomically distinct THY1^hiEGFR^hiPDGFRA^- bipotent glial progenitor cell (GPC), which is lineage-restricted to astrocytes and oligodendrocytes, but not to neurons. Our study provides a framework for the functional study of distinct cell types in human neurodevelopment.

Keywords: fluorescence-activated cell sorting; glial progenitor cells; index sorting; intermediate progenitor cells; neural stem cells; neurodevelopment; neuronal maturation; oligodendrocyte progenitor cells; radial glia; single-cell transcriptomics.