Efficient Induction of Oligodendrocyte Lineage From Pluripotent Stem Cells via Temporal Modulation of OLIG2 ( S147A ) and SOX10

Several protocols for generating oligodendrocytes (OLs) from human pluripotent stem cells have been reported. However, they are limited by long culture duration, intensive handling, and low yield of mature OLs. Transcription factor‐based strategies have improved efficiency, but OLIG2 and SOX10, key regulators of oligodendrocyte precursor cells (OPCs), also promote alternative neural fates. Here, we developed a Tet‐inducible system to control SOX10 and OLIG2 expression, including that of a phosphorylation‐deficient OLIG2 mutant (S147A). Co‐expression of SOX10 and OLIG2 enhanced OPC induction, confirmed by O4 positivity and transcriptomic profiling. Interestingly, only a brief induction of SOX10 + OLIG2(S147A) (2–5 days) efficiently yielded myelin basic protein positive OLs within 25 days, reaching approximately 20% of total cells. In contrast, sustained doxycycline‐mediated expression of SOX10 and OLIG2(S147A) favored OPC proliferation and delayed OL maturation. These findings highlight the importance of temporal control of transcription factor activity in accelerating OL differentiation and provide a practical platform for disease modeling and regenerative applications.