The Journal of Neuroscience, March 9, 2005, ():
Glioblastoma-Induced Attraction of Endogenous Neural Precursor Cells Is Associated with Improved Survival
J. Neurosci. Glass et al.
25: 2637
Supplemental data
Files in this Data Supplement:
- supplemental material
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supplemental-Figure-1. Characterization of G261-DsRed cells as invasive glioblastoma.
(a) G261-DsRed cells express GFAP in vivo. Photomultipliers of the confocal microscope were increased in sensitivity allowing the detection of GFAP (blue) in glioblastomas (arrow and center point of the 3-D reconstruction), but also providing a saturated signal for GFAP in nestin-GFP cells (arrowhead). In Fig. (3f) the sensitivity of the photomultiplier was set to a level allowing the detection of strong GFAP expression in parenchymal cells, but mainly excluding the weaker GFAP signal from glioblastoma cells. (b) In co-cultures of G261-DsRed cells with primary cultured astrocytes, strong GFAP expression (blue) is detected in the glia (arrowhead) and weaker, but nevertheless clear staining for GFAP is present in the glioblastoma (arrow). (c) Labeling for fibronectin (blue) is absent from glioblastomas and parenchymal nestin-GFP cells in vivo. (d) In co-cultures of fibroblasts with G261-DsRed cells, fibronectin (green) is abundant in fibroblast (arrowhead), but is only very weakly expressed in a minor fraction of glioblastoma cells (arrow). (e) Brevican (green) is expressed at the invasion front (arrows) of G261-DsRed glioblastoma cells (arrowhead). (f) Metalloproteinase-2 (MMP-2; blue) is expressed within G261-DsRed glioblastomas (arrowhead) and at the invasion front (arrows). (g) MMP-2 is active in G261 cultures; prominent bands for pro-MMP2 (72kDa) and MMP-2 (65kDa) are detected with gelatine-zymography.
Scales are: 30mm in (a to d) and 150mm in (e and f).
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supplemental-Figure-2. Characterization of nestin-GFP labeled cells accumulating around experimental glioblastomas.
(a) TUC-4, a marker for postmitotic neuronal precursors, is well detectable in the area close to the experimental glioblastoma and colabels with a subpopulation of nestin-GFP cells. (b) Calbindin, a marker for mature neurons, is expressed in the vicinity ofexperimental glioblastomas, but no significant colabeling with the population of nestin-gfp cells is noticeable. (c) Staining for the mature glia cell marker S-100? is restricted to the tumor area itself and a significant amount of colabeling with G261-dsred cells is evident. (d) Immunolabeling for CNP, a marker for oligodendrocyte precursors, colabeled with nestin-GFP in very few cells.
Scale bars of overviews are 25 µm.
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supplemental-Figure-3. Exogenously cultured neural precursors co-injected with glioblastoma cells reside at the glioblastoma and enwrap the tumor.
(a) Overview on a glioblastoma developing in a wild-type mouse 14 days after co-injection of nestin-GFP positive neural precursors together with G261-DsRed cells (mixing experiment); note that the precursors (green) surround the tumor mass (arrows). Detail of a mixing experiment, indicating that the precursors (green) enwrap invading glioblastoma cells (red).
Scales are: 120mm in (a) and 50mm in (b).
