Sox10-MCS5 enhancer dynamically tracks human oligodendrocyte progenitor fate
Autor: | Thomas R. Cimato, Fraser J. Sim, Melanie A. O'Bara, Jennifer K Lang, Andrew S. McCallion, Suyog Pol |
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Rok vydání: | 2013 |
Předmět: |
Receptor
Platelet-Derived Growth Factor alpha Cellular differentiation Green Fluorescent Proteins Induced Pluripotent Stem Cells Nerve Tissue Proteins Embryoid body Biology Article Green fluorescent protein OLIG2 Fetus Developmental Neuroscience medicine Humans Antigens Induced pluripotent stem cell Enhancer Cells Cultured Embryonic Stem Cells SOXE Transcription Factors O Antigens Cell Differentiation Flow Cytometry Molecular biology Embryonic stem cell Oligodendrocyte Oligodendroglia Enhancer Elements Genetic medicine.anatomical_structure Neurology embryonic structures Proteoglycans |
Zdroj: | Experimental Neurology. 247:694-702 |
ISSN: | 0014-4886 |
DOI: | 10.1016/j.expneurol.2013.03.010 |
Popis: | 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:GFPhigh 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. |
Databáze: | OpenAIRE |
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