SEZ cells populations

Characterization of neural stem cells (NSC) from the adult rodent subependymal niche by flow cytometry and traceable nucleoside retention assays have indicated the co-existence of activated and quiescent NSC in this niche. Single-cell deep-sequencing has further revealed different NSC molecular states with regards to cell-cycle gene expression: quiescent, primed for activation, and fully activated. Here, we provide a multi-level procedure to classify all cells of the subependymal niche, without the need for reporter mice, into populations with coherent transcriptomes that can be functionally assayed. Our strategy reveals that primed NSC are slowly proliferating cells that contribute to NSC cultures and that neurospheres contain NSC which retain a primed-like molecular signature and exhibit slow cycling and long-term self-renewal. Our strategy provides a way to functionally assay NSC heterogeneity and to study quiescence levels in NSC populations. Overall design: RNAseq of 8 different populations of cells freshly isolated from the adult mouse SEZ niche (between 3-4 replicates per group, 28 samples in total) and 2 populations of neurosphere forming cells showing different proliferative kinetics (4 replicates per group, 8 samples in total).