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

doi:10.1016/j.stemcr.2017.04.027

Stem Cell Reports (Jun 2017)

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

Affiliations

Gonçalo M.C. Rodrigues: Department of Bioengineering, University of California, Berkeley, Berkeley, CA 94720-1762, USA
Thomas Gaj: Department of Bioengineering, University of California, Berkeley, Berkeley, CA 94720-1762, USA
Maroof M. Adil: Department of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA 94720-1462, USA
Joyce Wahba: Department of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA 94720-1462, USA
Antara T. Rao: Department of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA 94720-1462, USA
Franziska K. Lorbeer: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720-3370, USA
Rishi U. Kulkarni: Department of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA 94720-1462, USA
Maria Margarida Diogo: Department of Bioengineering and Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, 1049-001 Lisbon, Portugal
Joaquim M.S. Cabral: Department of Bioengineering and Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, 1049-001 Lisbon, Portugal
Evan W. Miller: Department of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA 94720-1462, USA
Dirk Hockemeyer: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720-3370, USA
David V. Schaffer: Department of Bioengineering, University of California, Berkeley, Berkeley, CA 94720-1762, USA

DOI: https://doi.org/10.1016/j.stemcr.2017.04.027
Journal volume & issue: Vol. 8, no. 6
pp. 1770 – 1783

Abstract

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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.

Published in Stem Cell Reports

ISSN: 2213-6711 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Medicine: Medicine (General); Science: Biology (General)
Website: http://www.cell.com/stem-cell-reports/home

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