COBRE Journal Club:

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COBRE Journal Club

• Neural progenitors of the postnatal and adult
  mouse forebrain retain the ability to
  self-replicate, form neurospheres and undergo
  multipotent differentiation in vivo

Neumeister, B., Grabosch, A., Basak, O., Kemler,
R. and Taylor, V. Stem Cell Express, published
online 12/18/08
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Neural stem cell differentiation
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from Marci et al., J. Neurosci. 23(1)2003
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from Goldman, S., Nature Biotech. 232005
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from Rao et al., Anat. Rec. 2571999
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Do postnatal and adult EGF-dependent neurospheres
retain their primary stem cell feature to
self-replicate following long-term engraftment?
Experiment 1
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Figure 1A Schematic diagram of experimental
procedure

• isolated and expanded neurospheres from postnatal
  and adult mice that ubiquitously express
  bacterial ß-galactosidase or Renilla GFP
  (markers).
• using ultrasound backscatter microscopy,
  transplanted neurosphere-derived cells into
  forebrains of mice at mid-embryogenesis.

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Figure 1 Transplanted NSCs associate with the
host brain.
E 3 hours post transplantation.
F 24 hours post transplantation.
G 72 hours post transplantation.
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Can EGF-dependent neurospheres survive in the
host brain and differentiate?
Experiment 2
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Figure 2 Transplant-derived cells generate glia.
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Figure 3 Transplant- derived cells
generate neurons.
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Can transplanted cells retain the ability to
self-renew and generate neurospheresin vitro?
Experiment 3
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Figure 4 Re-transplantation of NSC showing
neurosphere self-renewal.
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Can transplant-derived neurospheres retain
neurospherogenic potential over multiple rounds
of transplantation?
Experiment 4
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Figure 4 Re-transplantation of NSC showing
neurosphere self-renewal.
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Figure 5 Re-transplanted NSCs generate
neurospheres that are multipotent.
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Are neurosphere-forming transplanted cells
mitotically active in vivo?
Experiment 5
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Figure 6 Transplanted NSCs proliferate in the
host brain.
E Most NSCs from primary hosts were generated
from BrdU cells.
F Most of the host-derived (GFP-) and
transplant-derived (GFP) neurospheres contain
BrdU label.
G Quantification of BrdU label-containing
neurospheres.
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Do mitotically active neurosphere forming
transplanted cells integrate into the neurogenic
brain regions of the host?
Experiment 6
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Figure 7 Transplanted NSCs proliferate in the
host brain.
Confocal image showing transplanted cells (GFP)
integrated into the subventricular zone (SVZ) of
the host brain.
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The End
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Figure X
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Figure X
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Neurosphere formation