Direct FACS Isolation of Neural Stem/Progenitor Lineages from the Adult Brain

Abstract

Adult neural stem and progenitor cells reside in the neurogenic niche of the adult brain and have tremendous potential in regenerative medicine. Compelling evidence suggests that adult neurogenesis plays an important role in hippocampal memory formation, plasticity, and mood regulation. Understanding the mechanisms that regulate the function of neural stem/progenitor cells within the brain is a critical step for the development of regenerative strategies to maintain or enhance neurological function. A major challenge in studying these cells is the limited cell number of adult neural stem cells, and the significant changes in their properties induced by in vitro culture and expansion. To best understand the regulation of these cells, they must be studied within their niche context. In this chapter, we provide a simplified protocol for the harvest and isolation of neural stem cell lineages directly from the murine brain, to provide input material for single-cell RNA-seq. This approach will elucidate the true transcriptional signatures and activated pathways in neural stem cell lineages, within the context of their niche environment.