Results: 5

1.
Fig. 2.

Fig. 2. From: CD133+ neural stem cells in the ependyma of mammalian postnatal forebrain.

Self-renewal capacity and multipotency of CD133+ ependymal cells. (a) A representative primary neurosphere at 6 DIV that originated clonally from a CD133+ ependymal cell. (Scale bar: 50 μm.) (b) Secondary spheres form upon dissociation of the primary neurospheres demonstrating the self-renewal capacity of CD133+ cells. (Scale bar: 50 μm.) (c) A representative primary sphere at 6 DIV originated clonally from a CD24+ ependymal cell. (Scale bar: 50 μm.) (d) Relative percentages of primary (pri.), secondary (sec.), tertiary (ter.), quaternary (qua.), and quinary (qui.) neurospheres originated from CD133+ ependymal cells. (e) The percentage of CD133+ ependymal cells in primary, secondary, and tertiary neurospheres at 7 DIV analyzed by FACS. (f) RT-PCR analysis of the expression of CD133, LeX, GFAP, and GAPDH from proliferating primary neurospheres at 6 DIV under bFGF treatment or from differentiated spheres in the absence of bFGF on coated slides for 7 days. (g–j) Fluorescent images taken from differentiated primary neurospheres originated from CD133+ ependymal cells (arrows in g) 3 or 7 (h–j) days after they were plated on coated slides after mitogen withdrawal. CD133+ neurospheres contain LeX+ progenitor/stem cells, TuJ1+ and MAP2+ neurons, CNPase+ oligodendrocytes, S100β+ and GFAP+ astrocytes, and nestin+ progenitor cells, demonstrating their multipotency. (Scale bars: g, 50 μm and h–i, 100 μm.)

Volkan Coskun, et al. Proc Natl Acad Sci U S A. 2008 January 22;105(3):1026-1031.
2.
Fig. 3.

Fig. 3. From: CD133+ neural stem cells in the ependyma of mammalian postnatal forebrain.

Proliferation and induction of CD133+ ependymal cells. (a and b) Occasional CD133+ ependymal cells (red) labeled with the cell proliferation marker Ki67 (green, arrowheads) analyzed with conventional (a) or confocal (b) microscopy. (Scale bars: a, 100 μm and b, 15 μm.) (c) CD133+ ependymal cells (red) incorporate BrdU (green) 14 days after the administration of Ara-C. Arrowheads point to BrdU+ cells in the ependymal layer. Arrows indicate BrdU+ cells in the SEZ. (Scale bar: 150 μm.) (d) A z-stack confocal analysis of a representative CD133+/BrdU+ cell (arrowhead). The arrow points to a BrdU+/CD133 cell in the SEZ. (Scale bar: 20 μm.) (e and f) Electron microscopic analysis of BrdU+ ependymal/subependymal cells upon AraC treatment. (e) Two BrdU ependymal cells (asterisks). (f) A BrdU+ ependymal cell (double asterisks) next to a BrdU ependymal cell (asterisk). Arrows mark BrdU+ cells in the SEZ. Note the similarities of the morphology of BrdU+ (double asterisks) and BrdU (asterisk) ependymal cells (e.g., shape of the nucleus, ciliated morphology, and round lipid droplets around the nuclei) and the differences between the BrdU+ ependymal cell (double asterisks) and subependymal cells (arrows). LV, lateral ventricle; SVZ, subventricular zone. (Scale bars: 4 μm.)

Volkan Coskun, et al. Proc Natl Acad Sci U S A. 2008 January 22;105(3):1026-1031.
3.
Fig. 1.

Fig. 1. From: CD133+ neural stem cells in the ependyma of mammalian postnatal forebrain.

Localization of CD133+ cells in the postnatal forebrain. (a and b) Sagittal sections of adult forebrain stained with CD133 (brown) and methyl green. The RMS is outlined with dotted lines. A subpopulation of ciliated ependymal cells lining the lateral ventricle is CD133+ (arrows in b). (Scale bars: a, 400 μm and b, 100 μm.) (c–e) Electron microscopic demonstration of the localization of CD133 to the membrane and the cilia of the ependymal cells. The section in c is not counterstained to demonstrate DAB depositions (arrows) on the cilia of the ependymal cells, whereas d shows a counterstained section to reveal subcellular structures. Note the characteristics of ependymal cells, such as lipid droplets, mirovilli, and cilia. Arrows in e indicate the gold particles that are concentrated on the cilia of ependymal cells. (Scale bars: c and d, 4 μm and e, 0.2 μm.) (f ) Detection of prominin-1 mRNA in the ependyma by in situ hybridization (arrows). (Scale bar: 100 μm.) (g) An adult forebrain section stained with CD133 (red) and the ciliar marker β-IV tubulin (green), showing CD133+/β-IV tubulin+ (yellow, arrows) and β-IV tubulin+/CD133 (green, arrowheads) ependymal cells. (Scale bar: 200 μm.) (h) Adult forebrain SVZ region immunostained with CD24 (green) and CD133 (red). Arrowheads indicate CD24+ ependymal cells in the ependyma and subependyma, whereas arrows point to CD133+ cells. (Upper Inset) CD133+ ependymal cells (arrows) and a CD24+ subependymal cell (arrowhead). (Lower Inset) CD24+ ependymal and subependymal cells (arrowheads) and CD24+/CD133+ ependymal cells (arrows). (i) An adult forebrain section stained with CD133 (red) and S100β (green), showing double-stained ependymal cells (arrows). (Scale bar: 200 μm). (j and k) Fluorescent photomicrographs of representative sagittal sections obtained by confocal microscopy from adult SVZ region. CD133+ ependymal cells (red) do not overlap with PSA-NCAM+ neuroblasts (green in j) or with GFAP+ cells (green in k) in the SEZ. The arrow in k points to GFAP+ processes touching the ventricle that are CD133. (Scale bars: 200 μm.) CC, corpus callosum; cp, choroid plexus; ld, lipid droplet; LV, lateral ventricle; Nu, nucleus; SVZ, subventricular zone.

Volkan Coskun, et al. Proc Natl Acad Sci U S A. 2008 January 22;105(3):1026-1031.
4.
Fig. 4.

Fig. 4. From: CD133+ neural stem cells in the ependyma of mammalian postnatal forebrain.

Contribution of transplanted CD133+ cells to the lineage of adult SVZ. (a) Confocal image of the SVZ region of adult Nestin/GFP mouse, stained with anti-CD133 (red). Arrows point to CD133+ ependymal cells. Arrowheads indicate Nestin-GFP cells in the SEZ. (Scale bar: 200 μm.) (b–e) Sorted CD133+ cells (red) from Nestin/GFP mice are not immunoreactive for GFP, LeX, GFAP, or Nestin during the first DIV. (Scale bars: b, 20 μm and c–e, 30 μm). (f–i) Clonally generated neurospheres from a CD133+ cell—isolated from Nestin/GFP mice—captured from the same area at 2 (f and g) or 6 (h and i) DIV with either bright field (f and h) or GFP (g and i) filters. Arrows point to Nestin/GFP+ spheres, whereas the arrowhead denotes a forming sphere with no GFP expression yet. (Scale bars: f and g, 75 μm and h and i, 100 μm.) (j and k) Differentiated spheres on coated surfaces at 5 DIV expressing Nestin/GFP and stained with LeX (red in j) or CD133 (red in k). (Scale bars: 100 μm.) (l) GFP+ cells (arrows) migrating along the RMS 2 weeks after transplantation of sorted CD133+/GFP cells from Nestin/GFP mice into the lateral ventricles of wild-type-recipient mice. (Scale bar: 250 μm.) (Insets) Higher magnification of migrating GFP+ cells in the RMS. (Scale bar: 20 μm.) (m) A confocal photomicrograph of the SVZ region of a recipient mouse 1 week after the sorted CD133+ cells were injected into the lateral ventricle. Even in close proximity of the ventricular surface, GFP+ cells (arrow) are CD133 (red). (n–r) Migrating GFP+ cells (green) in the SVZ/RMS are colabeled with doublecortin (arrowheads in n), GFAP (arrowhead in o), Mash1 (arrowhead in p) or PSA-NCAM (arrowheads in q). None of the GFP+ cells are costained with Olig2 (r). CC, corpus callosum; LV, lateral ventricle; RMS, rostral migratory stream; SVZ, subventricular zone. (Scale bars: n–o, 250 μm.)

Volkan Coskun, et al. Proc Natl Acad Sci U S A. 2008 January 22;105(3):1026-1031.
5.
Fig. 5.

Fig. 5. From: CD133+ neural stem cells in the ependyma of mammalian postnatal forebrain.

Generation of olfactory bulb interneurons by CD133+ ependymal neural stem cells in the postnatal forebrain. (a) GFP+ cells (green) that were transfected with an expression vector plasmid containing a short prominin-1 promoter (i.e., mP2) governing Cre-GFP expression line the ventricles (arrows). (Scale bar: 150 μm.) (b) A GFP+ transfected cell (arrow) in the ependymal layer is also CD133+ (red). (Scale bar: 15 μm.) (c and d) Fluorescent photomicrographs taken from Rosa26 reporter mice injected with mP2 plasmid. Transfected GFP+ cells (green) are in the ependyma whereas the β-gal+ cells (red) are in the ependyma and SEZ. Arrows point to transfected ependymal cells that started to express β-gal. (Scale bar: c, 200 μm and d, 50 μm.) (e) β-gal+ migrating cells (red) in the RMS are PSA-NCAM+ (green, arrowheads). (Scale bar: 75 μm.) (f) A migrating β-gal+ cell (red) in the layers of the olfactory bulb is TuJ1+ (green, arrowhead). Small arrowheads point to the process of the β-gal+ cell extending into the overlying layer of the olfactory bulb. (Scale bar: 50 μm.) (g) A photomontage of a P10 Rosa26 reporter mouse forebrain showing the entire extent of the RMS and olfactory bulb in a sagittal plane. The animal was injected with the mP2 construct at P3. β-gal+ cells (red) are distributed along the RMS from SVZ to the layers of the olfactory bulb. (Scale bar: 400 μm.) (h–k) Localization of β-gal+ cells in the RMS (arrows in h and i) and the layers of the olfactory bulb (arrows in j and k) of a P10 Rosa-26 reporter mouse. (Scale bars: 75 μm.) (l and m) A fluorescent photomicrograph of an adult Rosa26 reporter mouse forebrain injected with mP2 construct showing the SVZ region. β-gal+ cells (red) are distributed in the ependyma and SEZ, whereas transfected GFP+ cells line the ventricles (green, arrows). (Scale bar: 200 μm.) (n) Localization of β-gal+ cells (red) along the RMS en route to the olfactory bulb. Dotted white line outlines the borders of the RMS. (Scale bar: 250 μm.) (o) GFP+ transfected cells (green) in the ependymal layer of an adult Rosa26 reporter mouse are also CD133-immunoreactive (arrows, red). Hoechst staining (blue) was used to label nuclei. (Scale bar: 75 μm.) CC, corpus callosum; gcl, granule cell layer; gl, glomerular layer; LV, lateral ventricle; ml, mitral cell layer; OB, olfactory bulb; RMS, rostral migratory stream; SVZ, subventricular zone.

Volkan Coskun, et al. Proc Natl Acad Sci U S A. 2008 January 22;105(3):1026-1031.

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk