Charting the transition from in vitro production through in vivo diversification of human glial progenitor cells [CUT&Tag]

To better understand this process, we used scRNA-seq of >60,000 human cells to track the differentiation of hGPCs from pluripotent stem cells (PSCs), both in vitro and in vivo. In vitro, the hGPC pool comprised 4 transcriptionally-distinct subpopulations, the heterogeneity of which was associated with histone modifications of stage-dependent genes.

After the neonatal transplant of these progenitor cells into immunodeficient and myelin-deficient shiverer mice, they differentiated further; when extracted back from their host brains, scRNA-Seq revealed that the donor cells were comprised solely of hGPCs, astrocytes and oligodendrocytes. A combination of gene co-expression, motif enrichment, cell-trajectory, and cell-cell interaction analyses revealed that the tissue environment potentiated the further, context-dependent differentiation of hGPCs, via ligand-mediated activation of gene regulatory networks.