Identifying heterogeneity of Schwann cell like cells via scRNA-seq.

Previously we have reported that Schwann cell like cells (SCLCs) induced from human amniotic mesenchymal stem cells (hAMSCs) have better peripheral nerve regeneration ability, but the heterogeneity of SCLCs is still poorly understood. Here, based on single-cell RNA sequencing, we investigated the heterogeneity of hAMSCs and SCLCs, and identified the transition from hAMSCs to SCLCs. Totally 6,008 and 5,140 cells generated from hAMSCs and SCLCs were profiled and used for deep analysis. A subpopulation of SCLCs highly expressed Schwann cell markers, JUN, JUND, and NRG1, was identified. Then, cell cycle analysis distinguished the proliferation state of each cluster, and single cells of the subpopulation in SCLCs mainly remained in G0/G1 phase. Pseudotime analysis illustrated how hAMSCs transformed into SCLCs, hAMSCs showed more possibility transformed into other cell types. Through marker gene identification, pathway enrichment, transcription factor analysis, and secreted protein validation, genes such as NRG1, PTGS2, PITX1, VEGFA, and FGF2 were highly expressed in single cells. And also, pathways associated with nerve regeneration and immune control were enriched, such as angiogenesis or TNF signaling. The purpose of this study was to identify the heterogeneity of SCLCs. First, SCLCs was induced from hAMSCs. Then, the total RNA of hAMSCs and SCLCs was extracted for bulk RNA-seq(n=3); and also the RNA of single cells from hAMSCs and SCLCs was profiled via 10X genomics and bioinformatic analysis.