Abstract
We have developed a four-part protocol to differentiate human embryonic stem cells (hESCs) to oligodendrocyte progenitor cells (OPCs) according to developmental principles. In the first 2 weeks, hESCs are induced to differentiate into neuroepithelial cells, which form neural tube–like rosettes. In the following 10 d, these neuroepithelial cells are specified to OLIG2-expressing progenitors in the presence of retinoic acid (RA) and sonic hedgehog (SHH). Upon treatment with fibroblast growth factor 2 (FGF2) for another 10 d, these progenitors convert to OLIG2 and NKX2.2-expressing pre-OPCs. Finally, the pre-OPCs take 8–9 weeks to differentiate into OPCs, which express additional markers of oligodendrocytes, such as SOX10, platelet-derived growth factor receptor alpha (PDGFRα) and NG2. The unique aspects of the protocol are the use of FGF2 to promote the differentiation of gliogenic pre-OPCs in the third part and the removal of FGF2 during the transition of pre-OPCs to OPCs. This 3-month differentiation protocol consistently yields OPCs of high purity capable of producing myelin sheaths in vivo.
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Acknowledgements
This study was supported by the National Institute of Neurological Disorders and Stroke (R01 NS045926), the National Multiple Sclerosis Society (NMSS TR-3761), a gift from the Busta family and the Bleser family, and partly by a core grant to the Waisman Center from the National Institute of Child Health and Human Development (P30 HD03352).
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B.-Y.H. and Z.-W.D. designed and carried out experiments, analyzed data and wrote the paper; S.-C.Z. coordinated the study, designed experiments, analyzed data, wrote and finally approved the paper.
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Hu, BY., Du, ZW. & Zhang, SC. Differentiation of human oligodendrocytes from pluripotent stem cells. Nat Protoc 4, 1614–1622 (2009). https://doi.org/10.1038/nprot.2009.186
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DOI: https://doi.org/10.1038/nprot.2009.186
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