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Rapid and robust generation of functional oligodendrocyte progenitor cells from epiblast stem cells


Myelin-related disorders such as multiple sclerosis and leukodystrophies, for which restoration of oligodendrocyte function would be an effective treatment, are poised to benefit greatly from stem cell biology. Progress in myelin repair has been constrained by difficulties in generating pure populations of oligodendrocyte progenitor cells (OPCs) in sufficient quantities. Pluripotent stem cells theoretically provide an unlimited source of OPCs, but current differentiation strategies are poorly reproducible and generate heterogenous populations of cells. Here we provide a platform for the directed differentiation of pluripotent mouse epiblast stem cells (EpiSCs) through defined developmental transitions into a pure population of highly expandable OPCs in 10 d. These OPCs robustly differentiate into myelinating oligodendrocytes in vitro and in vivo. Our results demonstrate that mouse pluripotent stem cells provide a pure population of myelinogenic oligodendrocytes and offer a tractable platform for defining the molecular regulation of oligodendrocyte development and drug screening.

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Figure 1: Efficient differentiation of epiblast stem cells into region-specific neuroepithelial cells in 5 d.
Figure 2: Highly expandable OPCs derived from patterned EpiSC-derived neuroectoderm.
Figure 3: EpiSC-derived OPCs differentiated into oligodendrocytes in vitro.
Figure 4: EpiSC-derived OPCs are myelinogenic.
Figure 5: Screening for extrinsic signals that regulate the fate of EpiSC-derived OPCs.


  1. Goldman, S.A., Schanz, S. & Windrem, M.S. Stem cell-based strategies for treating pediatric disorders of myelin. Hum. Mol. Genet. 17, R1, R76–R83 (2008).

    Article  Google Scholar 

  2. Watkins, T.A., Emery, B., Mulinyawe, S. & Barres, B.A. Distinct stages of myelination regulated by γ-secretase and astrocytes in a rapidly myelinating CNS coculture system. Neuron 60, 555–569 (2008).

    Article  CAS  Google Scholar 

  3. Billon, N., Jolicoeur, C., Ying, Q.L., Smith, A. & Raff, M. Normal timing of oligodendrocyte development from genetically engineered, lineage-selectable mouse ES cells. J. Cell Sci. 115, 3657–3665 (2002).

    Article  CAS  Google Scholar 

  4. Brustle, O. et al. Embryonic stem cell-derived glial precursors: a source of myelinating transplants. Science 285, 754–756 (1999).

    Article  CAS  Google Scholar 

  5. Hu, B.Y., Du, Z.W. & Zhang, S.C. Differentiation of human oligodendrocytes from pluripotent stem cells. Nat. Protoc. 4, 1614–1622 (2009).

    Article  CAS  Google Scholar 

  6. Nistor, G.I., Totoiu, M.O., Haque, N., Carpenter, M.K. & Keirstead, H.S. Human embryonic stem cells differentiate into oligodendrocytes in high purity and myelinate after spinal cord transplantation. Glia 49, 385–396 (2005).

    Article  Google Scholar 

  7. Tam, P.P. & Loebel, D.A. Gene function in mouse embryogenesis: get set for gastrulation. Nat. Rev. Genet. 8, 368–381 (2007).

    Article  CAS  Google Scholar 

  8. Tesar, P.J. et al. New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature 448, 196–199 (2007).

    Article  CAS  Google Scholar 

  9. Brons, I.G. et al. Derivation of pluripotent epiblast stem cells from mammalian embryos. Nature 448, 191–195 (2007).

    Article  CAS  Google Scholar 

  10. Najm, F.J. et al. Isolation of epiblast stem cells from preimplantation mouse embryos. Cell Stem Cell 8, 318–325 (2011).

    Article  CAS  Google Scholar 

  11. Chambers, S.M. et al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat. Biotechnol. 27, 275–280 (2009).

    Article  CAS  Google Scholar 

  12. Hemmati-Brivanlou, A. & Melton, D.A. Inhibition of activin receptor signaling promotes neuralization in Xenopus. Cell 77, 273–281 (1994).

    Article  CAS  Google Scholar 

  13. Camus, A., Perea-Gomez, A., Moreau, A. & Collignon, J. Absence of Nodal signaling promotes precocious neural differentiation in the mouse embryo. Dev. Biol. 295, 743–755 (2006).

    Article  CAS  Google Scholar 

  14. Jessell, T.M. Neuronal specification in the spinal cord: inductive signals and transcriptional codes. Nat. Rev. Genet. 1, 20–29 (2000).

    Article  CAS  Google Scholar 

  15. Orentas, D.M. & Miller, R.H. The origin of spinal cord oligodendrocytes is dependent on local influences from the notochord. Dev. Biol. 177, 43–53 (1996).

    Article  CAS  Google Scholar 

  16. Trousse, F. et al. Notochord and floor plate stimulate oligodendrocyte differentiation in cultures of the chick dorsal neural tube. J. Neurosci. Res. 41, 552–560 (1995).

    Article  CAS  Google Scholar 

  17. Noll, E. & Miller, R.H. Oligodendrocyte precursors originate at the ventral ventricular zone dorsal to the ventral midline region in the embryonic rat spinal cord. Development 118, 563–573 (1993).

    CAS  PubMed  Google Scholar 

  18. Wichterle, H., Lieberam, I., Porter, J.A. & Jessell, T.M. Directed differentiation of embryonic stem cells into motor neurons. Cell 110, 385–397 (2002).

    Article  CAS  Google Scholar 

  19. Sun, T. et al. Olig bHLH proteins interact with homeodomain proteins to regulate cell fate acquisition in progenitors of the ventral neural tube. Curr. Biol. 11, 1413–1420 (2001).

    Article  CAS  Google Scholar 

  20. Krumlauf, R. et al. Hox homeobox genes and regionalisation of the nervous system. J. Neurobiol. 24, 1328–1340 (1993).

    Article  CAS  Google Scholar 

  21. Bogler, O., Wren, D., Barnett, S.C., Land, H. & Noble, M. Cooperation between two growth factors promotes extended self-renewal and inhibits differentiation of oligodendrocyte-type-2 astrocyte (o-2a) progenitor cells. Proc. Natl. Acad. Sci. USA 87, 6368–6372 (1990).

    Article  CAS  Google Scholar 

  22. Noble, M., Murray, K., Stroobant, P., Waterfield, M.D. & Riddle, P. Platelet-derived growth factor promotes division and motility and inhibits premature differentiation of the oligodendrocyte/type-2 astrocyte progenitor cell. Nature 333, 560–562 (1988).

    Article  CAS  Google Scholar 

  23. Richardson, W.D., Pringle, N.P., Yu, W.P. & Hall, A.C. Origins of spinal cord oligodendrocytes: Possible developmental and evolutionary relationships with motor neurons. Dev. Neurosci. 19, 58–68 (1997).

    Article  CAS  Google Scholar 

  24. Cahoy, J.D. et al. A transcriptome database for astrocytes, neurons, and oligodendrocytes: a new resource for understanding brain development and function. J. Neurosci. 28, 264–278 (2008).

    Article  CAS  Google Scholar 

  25. Barres, B.A., Lazar, M.A. & Raff, M.C. A novel role for thyroid hormone, glucocorticoids and retinoic acid in timing oligodendrocyte development. Development 120, 1097–1108 (1994).

    CAS  PubMed  Google Scholar 

  26. Chang, A., Nishiyama, A., Peterson, J., Prineas, J. & Trapp, B.D. NG2-positive oligodendrocyte progenitor cells in adult human brain and multiple sclerosis lesions. J. Neurosci. 20, 6404–6412 (2000).

    Article  CAS  Google Scholar 

  27. Fancy, S.P. et al. Dysregulation of the Wnt pathway inhibits timely myelination and remyelination in the mammalian CNS. Genes Dev. 23, 1571–1585 (2009).

    Article  CAS  Google Scholar 

  28. Hughes, S.M., Lillien, L.E., Raff, M.C., Rohrer, H. & Sendtner, M. Ciliary neurotrophic factor induces type-2 astrocyte differentiation in culture. Nature 335, 70–73 (1988).

    Article  CAS  Google Scholar 

  29. Mabie, P.C. et al. Bone morphogenetic proteins induce astroglial differentiation of oligodendroglial-astroglial progenitor cells. J. Neurosci. 17, 4112–4120 (1997).

    Article  CAS  Google Scholar 

  30. Windrem, M.S. et al. Neonatal chimerization with human glial progenitor cells can both remyelinate and rescue the otherwise lethally hypomyelinated shiverer mouse. Cell Stem Cell 2, 553–565 (2008).

    Article  CAS  Google Scholar 

  31. Chenoweth, J.G. & Tesar, P.J. Isolation and maintenance of mouse epiblast stem cells. Methods Mol. Biol. 636, 25–44 (2010).

    Article  Google Scholar 

  32. Mi, S. et al. Promotion of central nervous system remyelination by induced differentiation of oligodendrocyte precursor cells. Ann. Neurol. 65, 304–315 (2009).

    Article  CAS  Google Scholar 

  33. Schug, J. et al. Promoter features related to tissue specificity as measured by shannon entropy. Genome Biol. 6, R33 (2005).

    Article  Google Scholar 

  34. Cheadle, C., Vawter, M.P., Freed, W.J. & Becker, K.G. Analysis of microarray data using Z score transformation. J. Mol. Diagn. 5, 73–81 (2003).

    Article  CAS  Google Scholar 

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This research was supported by funding from US National Institutes of Health (MH087877 and NS30800), Case Western Reserve University School of Medicine, the Myelin Repair Foundation, the New York Stem Cell Foundation (Robertson Investigator award to P.J.T.), and the Cytometry and Imaging Microscopy, the Gene Expression and Genotyping, and the Radiation Resources Core facilities of the Case Comprehensive Cancer Center (P30 CA43703). S.N. was supported by the ENGAGE program of the Center for Stem Cell and Regenerative Medicine. We thank L. Cooperman, M. Hitomi, E. Hitomi, C. Batt, M. Pendergast, K. Wyatt, and members of the P. Scacheri and R. Atit laboratories for technical assistance and I. Tsung for artwork in Supplementary Figure 8, and J. Drazba, J. Peterson and K. Ryan for assistance with live-cell imaging.

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Authors and Affiliations



F.J.N. and P.J.T. derived the EpiSC-OPC lines and carried out in vitro experiments; A.V.C., A.Z., F.J.N. and P.J.T. performed in vivo experiments; A.Z., F.J.N., R.H.M. and P.J.T. performed slice culture experiments; E.C.F. carried out the co-culture experiments; F.J.N., S.N. and P.J.T. performed drug screening experiments; F.J.N., P.C.S. and P.J.T. generated and analyzed gene expression data; and F.J.N., R.H.M. and P.J.T. analyzed data and wrote the paper.

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Correspondence to Paul J Tesar.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–8, Supplementary Tables 1–2 (PDF 9057 kb)

Supplementary Video 1

Time-lapse imaging of EpiSC-derived OPCs differentiated to oligodendrocytes. Images were collected every 10 min for 68 h on an inverted microscope outfitted with a precision scanning stage in a live cell incubation chamber and cover at 37 °C and 5% CO2 in medium containing T3, SHH, noggin, cAMP, IGF1 and NT3. (MOV 8743 kb)

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Najm, F., Zaremba, A., Caprariello, A. et al. Rapid and robust generation of functional oligodendrocyte progenitor cells from epiblast stem cells. Nat Methods 8, 957–962 (2011).

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