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THE STEM CELL NICHE

Neural Stem Cells Express Non-Neural Markers During Embryoid Body Coculture

^a Centre for Reproduction and Development, Monash Institute of Medical Research, Clayton, Australia;
^b Department of Cytogenetics, Monash Medical Centre, Clayton, Australia;
^c Monash Immunology and Stem Cell Laboratories and
^d Department of Biochemistry and Molecular Biology Monash University, Clayton, Australia

Key Words. Embryonic stem cells • Neural stem cells • Smooth muscle • Cell fusion • Neural crest

Correspondence: Richard Mollard, Ph.D., Department of Biochemistry and Molecular Biology, Monash University, Clayton, 3800, Australia. Telephone: 61-3-9905-5753; Fax: 61-3-9905-3726; e-mail: mollard{at}med.monash.edu.au

Received April 5, 2005; accepted for publication October 13, 2005.
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 ß-galacosidase. Under these conditions, we assess neurosphere-derivative–immunoreactivity to anti-neurofilament heavy chain, anti-pan-cytokeratin, anti-smooth muscle -actinin and anti--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.