Isolation of adult rhesus neural stem and progenitor cells and differentiation into immature oligodendrocytes.
Neural stem and progenitor cells (NSPCs) have been isolated from several regions of the brain from mice, rats, and humans. These cells possess the characteristics of self-renewal and differentiation along all major neural lineages. Herein, the first isolation of NSPCs from the adult rhesus macaque brain and characterization of these cells based on their gene and protein expression profile, self-renewal, and ability to differentiate along an oligodendrocyte lineage are described. Flow cytometric analysis revealed that this cell population is CD90(+)/CD164(+)/CD34(-) and, therefore, resembles a nonhematopoietic stem cell population. Similar to other mesenchymal and neural stem cell populations, rhesus NSPCs cells express stemness-related genes, including the transcription factors Oct-4, Rex-1, and Sox-2 and the gene encoding for the intermediate filament protein nestin. The co-expression of the neural and glial markers MAP2ab, GFAP, NF-L, and NeuroD was also observed at both the mRNA and protein levels. When rhesus NSPCs were induced to differentiate with a cocktail of retinoic acid and the neurotrophins (NGF, BDNF, and NT-3), they underwent morphologic changes including taking on an oligodendrocyte precursor morphology. Along with these phenotypic changes, a decrease in MAP2ab expression and new expression of the oligodendrocyte precursor protein O4 were observed. Taken together, these results demonstrate the existence of a stem and progenitor cell-like population in the nonhuman primate brain, which may have the potential to generate oligodendroglia for use in the development of cellular therapies for demyelinating diseases.
Davis
Scott F
SF
Division of Gene Therapy, Tulane National Primate Center, Tulane University Health Sciences Center, Covington, LA 70433, USA.
Hood
Jay
J
Thomas
Anwyn
A
Bunnell
Bruce A
BA
D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D001921:Brain / Q000166:cytology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002469:Cell Separation; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics / Q000378:metabolism; D024002:HMGB Proteins / Q000235:genetics; D008253:Macaca mulatta; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D009414:Nerve Growth Factors / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D016900:Neurofilament Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D009836:Oligodendroglia / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055748:SOXB1 Transcription Factors; D013234:Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics; D014212:Tretinoin / Q000494:pharmacology
D015703:Antigens, CD; D051792:Basic Helix-Loop-Helix Proteins; D004268:DNA-Binding Proteins; D005904:Glial Fibrillary Acidic Protein; D024002:HMGB Proteins; D008869:Microtubule-Associated Proteins; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; D050814:Octamer Transcription Factor-3; D055748:SOXB1 Transcription Factors; D014157:Transcription Factors; D014212:Tretinoin; C527782:SOX2 protein, human; C527783:Sox2 protein, mouse; C527784:Sox2 protein, rat; C044882:neurofilament protein L; C093564:Neurogenic differentiation factor 1