IDENTIFICATION AND CHARACTERIZATION OF NEURAL STEM CELLS AND NEURAL PROGENITORS USING SINGLE-CELL ANALYSIS

Abstract

Neural stem cells (NSCs) are undifferentiated cells of the CNS which can self-renew and are multipotent. Neurospheres are used to propagate NSCs and neural progenitors (NPs). However, the exact cell types present within neurospheres is unknown. Furthermore, the identity of a neurosphere-forming cell (NFC) and NSC in vitro remains elusive. To identify cell types within neurospheres, I performed single-cell mRNA profiling of 48 genes in neurosphere cells. I identified three distinct cell populations which follow a developmental timeline of early, intermediate and late NPs. I then identified an NFC cluster and show that early NPs are the cell population that gives rise to neurospheres. Following which, I identified an NSC cluster, a bi/unipotent cluster and a non-NFC cluster which provide a clear distinction between NSCs and bi/unipotent NPs. Lastly, I derived the mRNA profile of a pure NSC population and identified candidate genes and signaling pathways important in NSC biology.