Journal: PLoS ONE
Article Title: Neuroprotective Effect of Transplanted Human Embryonic Stem Cell-Derived Neural Precursors in an Animal Model of Multiple Sclerosis
doi: 10.1371/journal.pone.0003145
Figure Lengend Snippet: (A–H) Immunofluorescence stainings of brain sections demonstrating the migration and differentiation of transplanted NPs, which were identified by the expression of human mitochondria (A, C–G), human nuclear antigens (B) and GFP (inset in A, H). (A): The NPs migrated extensively into white matter areas of the CNS such as the corpus callosum ( CC ) and were not observed in grey areas such as subcortical grey matter ( SGM ). Co-staining against the oligodendroglial marker, O4 (red) was used to identify the white matter. (B–H): Most of the transplanted cells either remained as uncommitted NPs expressing Musashi (B) or differentiated into early neuronal/oligodendroglial progenitors expressing Olig1 (C) or Olig2 (D). Further differentiation into more committed neuronal progenitors, oligodendrocyte progenitors or astrocytes expressing NGN2 (E), NG2 (F) and GFAP (G), respectively, was infrequent (∼1% of the transplanted cells per each of the cell types). Terminal differentiation of the transplanted NPs into GalC-expressing mature oligodendrocytes (H) was rare (<0.01% of the transplanted cells). Nuclei in A–H are counterstained with DAPI (blue). I–J: Toluidine blue stained transverse semi-thin sections of resin embedded spinal cords of NP-transplanted (J) and control (I) animals. In NP-transplanted animals, there were less demyelinated (arrows in I) and more normally myelinated axons than in controls. In both groups there were very rare remyelinated axons (G ratio>0.8). Scale bars: 15 µm (A–H), 5 µm (I–J).
Article Snippet: Immunofluorescent staining of NPs in vitro and in vivo: The following primary antibodies were used: Rabbit IgG anti-GFP (1∶100, Chemicon), mouse IgG anti-human specific mitochondria (1∶200, Chemicon), mouse IgM anti-A2B5 (1∶1, ATCC), mouse IgM anti-PSA-NCAM (1∶200, Chemicon), rabbit IgG anti-nestin (1∶50, Chemicon), rabbit anti-musashi (1∶100, Chemicon), mouse IgG anti-human nuclei (1∶50, Chemicon), rabbit IgG anti-NG2 (1∶50, Chemicon), mouse IgM anti-PDGFRα (1∶20, R&D), rabbit IgG anti-NGN2 (1∶300, Chemicon), rabbit anti-GalC (1∶20, Chemicon), mouse IgM anti-O4 (1∶20, Chemicon), rabbit IgG anti-MAP2 (1∶200, Chemicon), mouse IgG anti-β tubulin III (1∶2000, Sigma), rabbit anti-GFAP (1∶100, Dako), rabbit IgG anti-olig1 (1∶20, Chemicon) and goat anti-olig2 (1∶30, R&D).
Techniques: Immunofluorescence, Migration, Expressing, Staining, Marker
Proteintech is a verified supplier 
