title: Pluripotent stem cell-derived radial glia-like cells as stable intermediate for efficient generation of human oligodendrocytes.
abstract: Neural precursor cells (NPCs) derived from human pluripotent stem cells (hPSCs) represent an attractive tool for the in vitro generation of various neural cell types. However, the developmentally early NPCs emerging during hPSC differentiation typically show a strong propensity for neuronal differentiation, with more limited potential for generating astrocytes and, in particular, for generating oligodendrocytes. This phenomenon corresponds well to the consecutive and protracted generation of neurons and GLIA during normal human development. To obtain a more gliogenic NPC type, we combined growth factor-mediated expansion with pre-exposure to the differentiation-inducing agent retinoic acid and subsequent immunoisolation of CD133-positive cells. This protocol yields an adherent and self-renewing population of hindbrain/spinal cord radial glia (RG)-like neural precursor cells (RGL-NPCs) expressing typical neural stem cell markers such as nestin, ASCL1, SOX2, and PAX6 as well as RG markers BLBP, GLAST, vimentin, and GFAP. While RGL-NPCs maintain the ability for tripotential differentiation into neurons, astrocytes, and oligodendrocytes, they exhibit greatly enhanced propensity for oligodendrocyte generation. Under defined differentiation conditions promoting the expression of the major oligodendrocyte fate-determinants OLIG1/2, NKX6.2, NKX2.2, and SOX10, RGL-NPCs efficiently convert into NG2-positive oligodendroglial progenitor cells (OPCs) and are subsequently capable of in vivo myelination. Representing a stable intermediate between PSCs and OPCs, RGL-NPCs expedite the generation of PSC-derived oligodendrocytes with O4-, 4860-, and myelin basic protein (MBP)-positive cells that already appear within 7 weeks following growth factor withdrawal-induced differentiation. Thus, RGL-NPCs may serve as robust tool for time-efficient generation of human oligodendrocytes from embryonic and induced pluripotent stem cells.
authors.affiliation: Institute of Reconstructive Neurobiology, LIFE & BRAIN Center, University of Bonn, Germany.
authors.forename: Raphaela
authors.initials: R
authors.lastname: Gorris
authors.affiliation: Institute of Reconstructive Neurobiology, LIFE & BRAIN Center, University of Bonn, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Fischer
authors.affiliation: Institute of Reconstructive Neurobiology, LIFE & BRAIN Center, University of Bonn, Germany.
authors.forename: Kim Lina
authors.initials: KL
authors.lastname: Erwes
authors.affiliation: Institute of Reconstructive Neurobiology, LIFE & BRAIN Center, University of Bonn, Germany.
authors.forename: Jaideep
authors.initials: J
authors.lastname: Kesavan
authors.affiliation: Institute of Reconstructive Neurobiology, LIFE & BRAIN Center, University of Bonn, Germany.|Center for Stem Cell and Regenerative Medicine, Department of Neuroscience, Rosalind Franklin University of Medicine and Science, Chicago, Illinois.
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Peterson
authors.affiliation: Institute of Human Genetics, LIFE & BRAIN Center, University of Bonn, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Alexander
authors.affiliation: Institute of Human Genetics, LIFE & BRAIN Center, University of Bonn, Germany.
authors.forename: Markus M
authors.initials: MM
authors.lastname: Nöthen
authors.affiliation: Institute of Reconstructive Neurobiology, LIFE & BRAIN Center, University of Bonn, Germany.|German Center for Neurodegenerative Diseases, Bonn, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Peitz
authors.affiliation: Institute of Reconstructive Neurobiology, LIFE & BRAIN Center, University of Bonn, Germany.
authors.forename: Tamara
authors.initials: T
authors.lastname: Quandel
authors.affiliation: Institute of Reconstructive Neurobiology, LIFE & BRAIN Center, University of Bonn, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Karus
authors.affiliation: Institute of Reconstructive Neurobiology, LIFE & BRAIN Center, University of Bonn, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Brüstle
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D017690:Cell Transplantation; D053595:Embryonic Stem Cells / Q000502:physiology; D063928:Ependymoglial Cells / Q000502:physiology*; D005346:Fibroblast Growth Factors / Q000378:metabolism; D020869:Gene Expression Profiling; D006023:Glycoproteins / Q000378:metabolism; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins; D006801:Humans; D007150:Immunohistochemistry; D018345:Mice, Knockout; D008817:Mice, Mutant Strains; D009687:Nuclear Proteins; D009836:Oligodendroglia / Q000502:physiology*; D010455:Peptides / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors; D014212:Tretinoin / Q000378:metabolism
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins; C000715631:NKX2-2 protein, human; C000715627:Nkx2-2 protein, mouse; D009687:Nuclear Proteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D014157:Transcription Factors; D014212:Tretinoin; D005346:Fibroblast Growth Factors
keywords: gliogenesis; myelin; neural stem cells; radial glia cell