title: Generation and isolation of oligodendrocyte progenitor cells from human pluripotent stem cells.
abstract: In the CNS, oligodendrocytes act as the myelinating cells. Oligodendrocytes have been identified to be key players in several neurodegenerative disorders. This protocol describes a robust, fast and reproducible differentiation protocol to generate human oligodendrocytes from pluripotent stem cells (PSCs) using a chemically defined, growth factor-rich medium. Within 8 d, PSCs differentiate into paired box 6-positive (PAX6(+)) neural stem cells, which give rise to OLIG2(+) progenitors by day 12. Oligodendrocyte lineage transcription factor 2-positive (OLIG2(+)) cells begin to express the transcription factor NKX2.2 around day 18, followed by SRY-box 10 (SOX10) around day 40. Oligodendrocyte progenitor cells (OPCs) that are positive for the cell surface antigen recognized by the O4 antibody (O4(+)) appear around day 50 and reach, on average, 43% of the cell population after 75 d of differentiation. O4(+) OPCs can be isolated by cell sorting for myelination studies, or they can be terminally differentiated to myelin basic protein-positive (MBP(+)) oligodendrocytes. This protocol also describes an alternative strategy for markedly reducing the length and the costs of the differentiation and generating ∼30% O4(+) cells after only 55 d of culture.
authors.affiliation: The New York Stem Cell Foundation Research Institute, New York, New York, USA.
authors.forename: Panagiotis
authors.identifier: 000000024430997X
authors.initials: P
authors.lastname: Douvaras
authors.affiliation: The New York Stem Cell Foundation Research Institute, New York, New York, USA.
authors.forename: Valentina
authors.initials: V
authors.lastname: Fossati
mesh_terms: D002454:Cell Differentiation; D002469:Cell Separation; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins; D006801:Humans; D009687:Nuclear Proteins; D009836:Oligodendroglia / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000502:physiology*; D014157:Transcription Factors