Transcription factor-dependent differences in cluster composition and transition states of oligodendrocyte differentiation from human stem cells

Oligodendrocytes (OLs) are critical for myelination and are implicated in several brain disorders. Directed differentiation of human induced OLs (iOLs) from pluripotent stem cells can be achieved by forced expression of different combinations of the transcription factors SOX10 (S), OLIG2 (O) and NKX6.2 (N). Here, we applied quantitative image analysis and single-cell transcriptomics to compare different transcription factor combinations for their efficacy towards robust OL lineage conversion. Compared with S alone, the combination of SON significantly increases the number of myelin basic protein-positive iOLs and generates iOLs with a more complex morphology and higher expression levels of myelin-marker genes . The analysis of RNA velocity of individual cells reveals that S generates a population of volatile oligodendrocyte-precursor cells (OPCs) that appear to be more immature than those generated by SON and to display distinct molecular properties. Our analysis suggests that protocols for generating iOPCs or iOLs should be chosen depending on the intended application. Overall design: 3 scRNAseq samples in total: 1 hiPSC-derived NPC sample before induction of exogenous TF expression and 2 hiPSC-derived OL samples with different TF combinations after two weeks of differentiation