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The Journal of Neuroscience, November 15, 1999, 19(22):9986-9995
Identification, Isolation, and Promoter-Defined Separation of
Mitotic Oligodendrocyte Progenitor Cells from the Adult Human
Subcortical White Matter
Neeta Singh
Roy1, 3,
Su
Wang1, 3,
Catherine
Harrison-Restelli1,
Abdellatif
Benraiss1,
Richard A. R.
Fraser2,
Michel
Gravel4,
Peter E.
Braun4, and
Steven A.
Goldman1
Departments of 1 Neurology and Neuroscience and
2 Neurosurgery, Cornell University Medical College, New
York, New York 10021, 3 Aitken Neuroscience Center, New
York, New York 10021, and 4 Department of Biochemistry,
McGill University, Montreal, Quebec, Canada H3A 2T5
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.
Key words:
regeneration; myelin; remyelination; cell sorting; stem
cells; subependyma
Copyright © 1999 Society for Neuroscience 0270-6474/99/19229986-10$05.00/0
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