Defined and Scalable Differentiation of Human Oligodendrocyte Precursors from Pluripotent Stem Cells in a 3D Culture System

Gonçalo M.C. Rodrigues, Thomas Gaj, Maroof M. Adil, Joyce Wahba, Antara T. Rao, Franziska K. Lorbeer, Rishi U. Kulkarni, Maria Margarida Diogo, Joaquim M.S. Cabral, Evan W. Miller, Dirk Hockemeyer, David V. Schaffer

Research output: Contribution to journalArticle

Abstract

Oligodendrocyte precursor cells (OPCs) offer considerable potential for the treatment of demyelinating diseases and injuries of the CNS. However, generating large quantities of high-quality OPCs remains a substantial challenge that impedes their therapeutic application. Here, we show that OPCs can be generated from human pluripotent stem cells (hPSCs) in a three-dimensional (3D), scalable, and fully defined thermoresponsive biomaterial system. We used CRISPR/Cas9 to create a NKX2.2-EGFP human embryonic stem cell reporter line that enabled fine-tuning of early OPC specification and identification of conditions that markedly increased the number of OLIG2+ and NKX2.2+ cells generated from hPSCs. Transplantation of 50-day-old OPCs into the brains of NOD/SCID mice revealed that progenitors generated in 3D without cell selection or purification subsequently engrafted, migrated, and matured into myelinating oligodendrocytes in vivo. These results demonstrate the potential of harnessing lineage reporter lines to develop 3D platforms for rapid and large-scale production of OPCs.

Original languageEnglish (US)
Pages (from-to)1770-1783
Number of pages14
JournalStem Cell Reports
Volume8
Issue number6
DOIs
StatePublished - Jun 6 2017

Keywords

  • CRISPR/Cas9 knockin
  • NKX2.2-EGFP reporter cell line
  • human pluripotent stem cells
  • oligodendrocyte precursor cells
  • scalable and defined oligodendrocyte differentiation
  • three-dimensional culture systems

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Developmental Biology
  • Cell Biology

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  • Cite this

    Rodrigues, G. M. C., Gaj, T., Adil, M. M., Wahba, J., Rao, A. T., Lorbeer, F. K., Kulkarni, R. U., Diogo, M. M., Cabral, J. M. S., Miller, E. W., Hockemeyer, D., & Schaffer, D. V. (2017). Defined and Scalable Differentiation of Human Oligodendrocyte Precursors from Pluripotent Stem Cells in a 3D Culture System. Stem Cell Reports, 8(6), 1770-1783. https://doi.org/10.1016/j.stemcr.2017.04.027