Sox10-MCS5 enhancer dynamically tracks human oligodendrocyte progenitor fate

Suyog U. Pol, Jennifer K. Lang, Melanie A. O'Bara, Thomas R. Cimato, Andrew S McCallion, Fraser J. Sim

Research output: Contribution to journalArticle

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: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.

Original languageEnglish (US)
Pages (from-to)694-702
Number of pages9
JournalExperimental Neurology
Volume247
DOIs
StatePublished - Sep 2013

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Keywords

  • Enhancer
  • Lentivirus
  • Pluripotent
  • SOX10

ASJC Scopus subject areas

  • Neurology
  • Developmental Neuroscience

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