Identification, isolation, and promoter-defined separation of mitotic oligodendrocyte progenitor cells from the adult human subcortical white matter

Neeta Singh Roy, Su Wang, Catherine Harrison-Restelli, Abdellatif Benraiss, Richard A.R. Fraser, Michel Gravel, Peter E. Braun, Steven A. Goldman

Research output: Contribution to journalArticlepeer-review

212 Scopus citations

Abstract

Previous studies have suggested the persistence of oligodendrocyte progenitor cells in the adult mammalian subcortical white matter. To identify oligodendrocyte progenitors in the adult human subcortical white matter, we transfected dissociates of capsular white matter with plasmid DNA bearing the gene for green fluorescence protein (hGFP), placed under the control of the human early promoter (P2) for the oligodendrocytic protein cyclic nucleotide phosphodiesterase (P/hCNP2). Within 4 d after transfection with P/hCNP2:hGFP, a discrete population of small, bipolar cells were noted to express GFP. These cells were A2B5-positive (A2B5+), incorporated bromodeoxyuridine in vitro, and constituted <0.5% of all cells. Using fluorescence-activated cell sorting (FACS), the P/hCNP2-driven GFP- cells were then isolated and enriched to near-purity. In the weeks after FACS, most P/hCNP2:hGFP-sorted cells matured as morphologically and antigenically characteristic oligodendrocytes. Thus, the human subcortical white matter harbors mitotically competent progenitor cells, which give rise primarily to oligodendrocytes in vitro. By using fluorescent transgenes of GFP expressed under the control of an early oligodendrocytic promoter, these oligodendrocyte progenitor cells may be extracted and purified from adult human white matter in sufficient numbers for implantation and cell-based therapy.

Original languageEnglish
Pages (from-to)9986-9995
Number of pages10
JournalJournal of Neuroscience
Volume19
Issue number22
DOIs
StatePublished - 15 Nov 1999
Externally publishedYes

Keywords

  • Cell sorting
  • Myelin
  • Regeneration
  • Remyelination
  • Stem cells
  • Subependyma

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