抄録
For isolation neural stem cells (NSCs), long-term in vitro cultivation is necessary because of the lack of valid cell surface markers suitable for prospective purification, although long term cultivation may lead to phenotype deterioration. Here, we focused on oligosaccharide chains on NPC surface as a valid marker for isolation. Fluorescence-activated cell sorting (FACS) analysis and immunohistochemical analysis revealed the expression of biantennary complex type N-glycans that recognized by erythroagglutinating-Phaseolus vulgaris agglutinin (E-PHA) lectin in nestin-positive cells in the neurogenic area. In addition, E-PHA bound preferentially to purified NSCs as well as to astrocytes, but not to neurons, microglia, and oligodendrocyte precursor cells. Such high affinity E-PHA binding to NSCs prompted us to use E-PHA as a cell surface marker for NSC isolation by using FACS. In E-PHA binding cells, we found about 30-fold increase in the number of neurosphere-forming cells, capable of differentiating into mature neurons or glial cells, compared to that of non-sorting cells. This indicates that E-PHA, which recognizes complex-type N-glycans, is a valid cell surface marker of mouse NSCs for prospective isolation. [J Physiol Sci. 2008;58 Suppl:S97]
