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

Nature Methods volume 8, pages 957–962 (2011)Cite this article

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Abstract

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.

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Acknowledgements

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

  1. Department of Genetics, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA

    Fadi J Najm, Shreya Nayak, Peter C Scacheri & Paul J Tesar

  2. Department of Neurosciences, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA

    Anita Zaremba, Andrew V Caprariello, Robert H Miller & Paul J Tesar

  3. Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, USA

    Eric C Freundt

  4. New York Stem Cell Foundation, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA

    Paul J Tesar

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  1. Fadi J Najm
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Contributions

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). https://doi.org/10.1038/nmeth.1712

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