biostudies-literatureLi WNEI NIH HHSNINDS NIH HHS106156https://www.ebi.ac.uk/biostudies/studies/S-EPMC9958491biostudies-literatureUnknown26(3)Promoting myelination capacity of endogenous oligodendrocyte precursor cells (OPCs) is a promising therapeutic approach for CNS demyelinating disorders such as Multiple Sclerosis (MS). To aid in the discovery of myelination-promoting compounds, we generated a genome-engineered human pluripotent stem cell (hPSC) line that consists of three reporters: identification-and-purification tag, GFP, and secreted-NanoLuc, driven by the endogenous <i>PDGFRA, PLP1,</i> and <i>MBP</i> genes, respectively. Using this cell line, we established a high-throughput drug screening platform and performed a small-molecule screen, which identified at least two myelination-promoting small-molecule (Ro1138452 and SR2211) that target prostacyclin (IP) receptor and retinoic acid receptor-related orphan receptor γ (RiScienceHigh-throughput screening for myelination promoting compounds using human stem cell-derived oligodendrocyte progenitor cells.PMC9958491R00 EY029011P30 EY001765R21 NS096668K99 EY029011Dodge JCMao HQJi HTakaesu FBerlinicke CChen CZack DJGiera SChang XLi WKumar DSheng GMcGrath AGHuang YFang WChamling XChang CMadden SDuan YfalseHigh-throughput screening for myelination promoting compounds using human stem cell-derived oligodendrocyte progenitor cells.Promoting myelination capacity of endogenous oligodendrocyte precursor cells (OPCs) is a promising therapeutic approach for CNS demyelinating disorders such as Multiple Sclerosis (MS). To aid in the discovery of myelination-promoting compounds, we generated a genome-engineered human pluripotent stem cell (hPSC) line that consists of three reporters: identification-and-purification tag, GFP, and secreted-NanoLuc, driven by the endogenous <i>PDGFRA, PLP1,</i> and <i>MBP</i> genes, respectively. Using this cell line, we established a high-throughput drug screening platform and performed a small-molecule screen, which identified at least two myelination-promoting small-molecule (Ro1138452 and SR2211) that target prostacyclin (IP) receptor and retinoic acid receptor-related orphan receptor γ (R2023-01-01T00:00:00Z2023 Mar2026-05-29T05:30:08.18Z2025-02-18T23:44:44.386ZS-EPMC99584913685228110.1016/j.isci.2023.106156