title: Sox10-MCS5 enhancer dynamically tracks human oligodendrocyte progenitor fate.
abstract: In this study, we sought to establish a novel method to prospectively and dynamically identify live human oligodendrocyte precursor cells (OPCs) and oligodendrocyte lineage cells from brain dissociates and pluripotent stem cell culture. We selected a highly conserved enhancer element of the Sox10 gene, known as MCS5, which directs reporter expression to oligodendrocyte lineage cells in mouse and zebrafish. We demonstrate that lentiviral Sox10-MCS5 induced expression of GFP at high levels in a subpopulation of human CD140a/PDGFαR-sorted OPCs as well as their immature oligodendrocyte progeny. Furthermore, we show that almost all Sox10-MCS5:GFP(high) cells expressed OPC antigen CD140a and human OPCs expressing SOX10, OLIG2, and PDGFRA mRNAs could be prospectively identified using GFP based fluorescence activated cells sorting alone. Additionally, we established a human induced pluripotent cell (iPSC) line transduced with the Sox10-MCS5:GFP reporter using a Rex-Neo cassette. Similar to human primary cells, GFP expression was restricted to embryoid bodies containing both oligodendrocyte progenitor and oligodendrocyte cells and co-localized with NG2 and O4-positive cells respectively. As such, we have developed a novel reporter system that can track oligodendrocyte commitment in human cells, establishing a valuable tool to improve our understanding and efficiency of human oligodendrocyte derivation.
authors.affiliation: Department of Pharmacology, School of Medicine and Biomedical Sciences, University at Buffalo, Buffalo, NY 14214, USA.
authors.forename: Suyog U
authors.initials: SU
authors.lastname: Pol
authors.forename: Jennifer K
authors.initials: JK
authors.lastname: Lang
authors.forename: Melanie A
authors.initials: MA
authors.lastname: O'Bara
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Cimato
authors.forename: Andrew S
authors.initials: AS
authors.lastname: McCallion
authors.forename: Fraser J
authors.initials: FJ
authors.lastname: Sim
mesh_terms: D000941:Antigens / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000502:physiology*; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D005333:Fetus; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D019081:O Antigens / Q000378:metabolism; D009836:Oligodendroglia / Q000378:metabolism*; D011509:Proteoglycans / Q000378:metabolism; D020796:Receptor, Platelet-Derived Growth Factor alpha / Q000378:metabolism; D055757:SOXE Transcription Factors / Q000235:genetics / Q000378:metabolism*; D000099216:Chondroitin Sulfate Proteoglycan 4
chemical_list: D000941:Antigens; D049452:Green Fluorescent Proteins; D009419:Nerve Tissue Proteins; D019081:O Antigens; D011509:Proteoglycans; D020796:Receptor, Platelet-Derived Growth Factor alpha; D055757:SOXE Transcription Factors; D000099216:Chondroitin Sulfate Proteoglycan 4; C527923:SOX10 protein, human
keywords: Enhancer; Lentivirus; Pluripotent; SOX10