Efficient Generation of Viral and Integration-Free Human Induced Pluripotent Stem Cell-Derived Oligodendrocytes

Araceli Espinosa-Jeffrey; Bruno Blanchi; Juan Carlos Biancotti; Shalini Kumar; Megumi Hirose; Berhan Mandefro; Dodanim Talavera-Adame; Nissim Benvenisty; Jean de Vellis

doi:10.1002/cpsc.19

Efficient Generation of Viral and Integration-Free Human Induced Pluripotent Stem Cell-Derived Oligodendrocytes

Curr Protoc Stem Cell Biol. 2016 Nov;39(1):2D.18.1-2D.18.28. doi: 10.1002/cpsc.19.

Authors

Araceli Espinosa-Jeffrey¹, Bruno Blanchi¹, Juan Carlos Biancotti², Shalini Kumar¹, Megumi Hirose¹, Berhan Mandefro³, Dodanim Talavera-Adame³, Nissim Benvenisty⁴, Jean de Vellis¹

Affiliations

¹ Intellectual and Developmental Disabilities Research Center, Semel Institute for Neuroscience, David Geffen School of Medicine at UCLA, Los Angeles, California.
² Zilkha Neurogenetic Institute, Keck School of Medicine, University of Southern California, Los Angeles, California.
³ Regenerative Medicine Institute, Cedars Sinai Medical Center, Los Angeles, California.
⁴ Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel.

PMID: 31816188
DOI: 10.1002/cpsc.19

Abstract

Here we document three highly reproducible protocols: (1) a culture system for the derivation of human oligodendrocytes (OLs) from human induced pluripotent stem cells (hiPS) and their further maturation-our protocol generates viral- and integration-free OLs that efficiently commit and move forward in the OL lineage, recapitulating all the steps known to occur during in vivo development; (2) a method for the isolation, propagation and maintenance of neural stem cells (NSCs); and (3) a protocol for the production, isolation, and maintenance of OLs from perinatal rodent and human brain-derived NSCs. Our unique culture systems rely on a series of chemically defined media, specifically designed and carefully characterized for each developmental stage of OL as they advance from OL progenitors to mature, myelinating cells. We are confident that these protocols bring our field a step closer to efficient autologous cell replacement therapies and disease modeling. © 2016 by John Wiley & Sons, Inc.

Keywords: NSC; chemically defined media; human induced pluripotent stem cells; lineage progression; neural stem cells; neurospheres; oligodendrocyte maturation; oligodendrocyte specification; oligospheres.