Identifying Genes that Affect Differentiation of Human Neural Stem Cells and Myelination of Mature Oligodendrocytes

Abstract

Human neural stem cells (NSCs) are self-renewing, multipotent cells of the central nervous system (CNS). They are characterized by their ability to differentiate into a range of cells, including oligodendrocytes (OLs), neurons, and astrocytes, depending on exogenous stimuli. An efficient and easy directional differentiation method was developed for obtaining large quantities of high-quality of human OL progenitor cells (OPCs) and OLs from NSCs. RNA sequencing, immunofluorescence staining, flow cytometry, western blot, label-free proteomic sequencing, and qPCR were performed in OL lines differentiated from NSC lines. The changes in the positive rate of typical proteins were analyzed expressed by NSCs, neurons, astrocytes, OPCs, and OLs. We assessed Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of differentially expressed (DE) messenger RNAs (mRNAs) related to the differentiation of NSCs and the maturation of OLs. The percentage of NSCs differentiated into neurons, astrocytes, and OLs was 82.13%, 80.19%, and 90.15%, respectively. We found that nestin, PAX6, Musashi, and vimentin were highly expressed in NSCs; PDGFR-α, A2B5, NG2, OLIG2, SOX10, and NKX2-2 were highly expressed in OPCs; and CNP, GALC, PLP1, and MBP were highly expressed in OLs. RNA sequencing, western blot and qPCR revealed that ERBB4 and SORL1 gradually increased during NSC–OL differentiation. In conclusion, NSCs can differentiate into neurons, astrocytes, and OLs efficiently. PDGFR-α, APC, ID4, PLLP, and other markers were related to NSC differentiation and OL maturation. Moreover, we refined a screening method for ERBB4 and SORL1, which may underlie NSC differentiation and OL maturation. Graphical Abstract Potential unreported genes and proteins may regulate differentiation of human neural stem cells into oligodendrocyte lineage. Neural stem cells (NSCs) can differentiate into neurons, astrocytes, and oligodendrocyte (OLs) efficiently. By analyzing the DE mRNAs and proteins of NSCs and OLs lineage, we could identify reported markers and unreported markers of ERBB4 and SORL1 that may underlie regulate NSC differentiation and OL maturation.

Synergistic Effect of Electrical and Biochemical Stimulation on Human iPSC-Derived Neural Differentiation in a Microfluidic Electrode Array Chip

“…In contrast, the NF+ES group showed a higher prevalence of cells expressing differentiated Olig2 (green signal). During nervous system development, there is a stage where Olig2 and nestin are coexpressed in certain neural precursor cells, representing a transitional state during their differentiation into oligodendrocytes . Furthermore, nestin is known to be expressed in early stages of neural development, including oligodendrocyte precursor cells (OPCs), and is gradually downregulated as OPCs differentiate into mature oligodendrocytes. , Therefore, the increased level of nestin expression in the single stimulus group suggests a transitional state of differentiation, indicating the presence of progenitor cells that have not fully differentiated into oligodendrocytes or glial cells.…”

Neuroinflammation Induces Myelin Damage by Inhibiting Gapdh of Oligodendrocytes in the Hippocampus

“…Cluster 0 was characterized by markers of mature oligodendrocytes, including Ptgds [53], and genes associated with myelin sheath and oligodendrocyte maturation, such as Mal, Olig1, Pllp [54,52,11], and Htra1 [55]. Functional enrichment analysis suggested that this cluster represented mature oligodendrocytes involved in myelination.…”

The roles of neural stem cells in myelin regeneration and repair therapy after spinal cord injury

“…They have indefinite self-renewal capacity and the ability to give rise to neural progenitor cells (NPCs). Additionally, they have the potential to continue developing into mature neural lineages, such as neurons, astrocytes and oligodendrocytes [ 35 – 37 ].…”