Cell surface N-glycans mediated isolation of mouse neural stem cells
Article first published online: 1 JUL 2009
© 2009 The Authors. Journal Compilation © 2009 International Society for Neurochemistry
Journal of Neurochemistry
Volume 110, Issue 5, pages 1575–1584, September 2009
How to Cite
Hamanoue, M., Matsuzaki, Y., Sato, K.-i., Okano, H. J., Shibata, S., Sato, I., Suzuki, S., Ogawara, M., Takamatsu, K. and Okano, H. (2009), Cell surface N-glycans mediated isolation of mouse neural stem cells. Journal of Neurochemistry, 110: 1575–1584. doi: 10.1111/j.1471-4159.2009.06256.x
- Issue published online: 10 AUG 2009
- Article first published online: 1 JUL 2009
- Received February 19, 2009; revised manuscript received June 6, 2009; accepted June 17, 2009.
- cell separation;
- flow cytometry;
- membrane glycoproteins;
- plant lectins;
- stem cells
The isolation of neural stem cells (NSCs) from the brain has been hampered by the lack of valid cell surface markers and the requirement for long-term in vitro cultivation that may lead to phenotype deterioration. However, few suitable specific cell surface antigens are available on NSCs that could be used for their prospective isolation. The present study demonstrated that the expression of complex type asparagine-linked oligosaccharide (N-glycans) was detected on brain cells dissociated from embryonic and adult brain using Phaseolus vulgaris erythroagglutinating lectin (E-PHA) which binds to biantennary complex type N-glycans, and demonstrated that E-PHA bound preferentially to purified NSCs, but not to neurons, microglia, or oligodendrocyte precursor cells. The labeling of dissociated mouse embryonic brain cells or adult brain cells with E-PHA enabled the enrichment of NSCs by 25-fold or 9-fold of the number of neurosphere-forming cells in comparison to that of unsorted cells, respectively. Furthermore, a lectin blot analysis revealed the presence of several glycoproteins which were recognized by E-PHA in the membrane fraction of the proliferating NSCs, but not in the differentiated cells. These results indicate that complex type N-glycans is a valuable cell surface marker for living mouse NSCs from both the embryonic and adult brain.