title: Differentiation of embryonic stem cells into oligodendrocyte precursors.
abstract: Oligodendrocytes are the myelinating cells of the central nervous system. For regenerative cell therapy in demyelinating diseases, there is significant interest in deriving a pure population of lineage-committed oligodendrocyte precursor cells (OPCs) for transplantation. OPCs are characterized by the activity of the transcription factor Olig2 and surface expression of a proteoglycan NG2. Using the GFP-Olig2 (G-Olig2) mouse embryonic stem cell (mESC) reporter line, we optimized conditions for the differentiation of mESCs into GFP+Olig2+NG2+ OPCs. In our protocol, we first describe the generation of embryoid bodies (EBs) from mESCs. Second, we describe treatment of mESC-derived EBs with small molecules: (1) retinoic acid (RA) and (2) a sonic hedgehog (Shh) agonist purmorphamine (Pur) under defined culture conditions to direct EB differentiation into the oligodendroglial lineage. By this approach, OPCs can be obtained with high efficiency (>80%) in a time period of 30 days. Cells derived from mESCs in this protocol are phenotypically similar to OPCs derived from primary tissue culture. The mESC-derived OPCs do not show the spiking property described for a subpopulation of brain OPCs in situ. To study this electrophysiological property, we describe the generation of spiking mESC-derived OPCs by ectopically expressing Na(V;)1.2 subunit. The spiking and nonspiking cells obtained from this protocol will help advance functional studies on the two subpopulations of OPCs.
authors.affiliation: Department of Cell Biology and Human Anatomy Institute for Pediatric Regenerative Medicine, School of Medicine, University of California, Davis, CA, USA.
authors.forename: Peng
authors.initials: P
authors.lastname: Jiang
authors.forename: Vimal
authors.initials: V
authors.lastname: Selvaraj
authors.forename: Wenbin
authors.initials: W
authors.lastname: Deng
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D003584:Cytological Techniques / Q000379:methods*; D055724:Electrophysiological Phenomena; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D009025:Morpholines / Q000494:pharmacology; D009836:Oligodendroglia / Q000166:cytology* / Q000187:drug effects; D011687:Purines / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D009025:Morpholines; D011687:Purines; D014212:Tretinoin; C470893:purmorphamine