Neural stem cells express non-neural markers during embryoid body coculture.
The capacity of neural stem cells (NSC) to transdifferentiate into a wide range of non-neuronal lineages is the subject of debate. One approach to test NSC plasticity is to ectopically place NSCs in permissive or instructive microenvironments in which the signals driving differentiation of multiple cell types are being elicited. Here we produce embryoid body neurosphere aggregates by combining neurosphere derivatives from fetal mice constitutively expressing green fluorescent protein with embryonic stem (ES) cells isolated from Zin40 mice constitutively expressing nuclear beta-galacosidase. Under these conditions, we assess neurosphere-derivative-immunoreactivity to anti-neurofilament heavy chain, anti-pan-cytokeratin, anti-smooth muscle alpha-actinin and anti-alpha-fetoprotein-specific antibodies. Furthermore, we determine lineage-specific transgene expression and undertake fluorescence in situ hybridization to assess ES cell-neural stem cell-fusion indices. Our data demonstrate that following coculture in hanging drops with ES cells, neurosphere derivatives display immunoreactivity to non-neural markers, in particular smooth muscle, which is not dependent upon cell-cell fusion. These results suggest that given an appropriate environment, NSC may lose their in vivo restrictions and display non-neuronal phenotypes.
Denham
Mark
M
Centre for Reproduction and Development, Monash Institute of Medical Research, Clayton, Australia.
Huynh
Trieu
T
Dottori
Mirella
M
Allen
Greg
G
Trounson
Alan
A
Mollard
Richard
R
D000818:Animals; D001483:Base Sequence; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics; D015415:Biomarkers; D001921:Brain / Q000166:cytology; D002454:Cell Differentiation; D002460:Cell Line; D002469:Cell Separation; D002678:Chimera; D018920:Coculture Techniques; D004247:DNA / Q000235:genetics; D005333:Fetus / Q000166:cytology; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
D051792:Basic Helix-Loop-Helix Proteins; D015415:Biomarkers; D004247:DNA; D049452:Green Fluorescent Proteins; D011994:Recombinant Proteins; C499317:Ascl1 protein, mouse