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Exp Neurol. 2013 Sep;247:694-702. doi: 10.1016/j.expneurol.2013.03.010. Epub 2013 Mar 16.

Sox10-MCS5 enhancer dynamically tracks human oligodendrocyte progenitor fate.

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  • 1Department of Pharmacology, School of Medicine and Biomedical Sciences, University at Buffalo, Buffalo, NY 14214, USA.

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.

Copyright © 2013 Elsevier Inc. All rights reserved.

KEYWORDS:

Enhancer; Lentivirus; Pluripotent; SOX10

PMID:
23507034
[PubMed - indexed for MEDLINE]
PMCID:
PMC4254711
Free PMC Article
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