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Cell surface N‐ glycans mediated isolation of mouse neural stem cells
Author(s) -
Hamanoue Makoto,
Matsuzaki Yumi,
Sato Kenichiro,
Okano Hirotaka James,
Shibata Shinsuke,
Sato Isamu,
Suzuki Sadafumi,
Ogawara Miyuki,
Takamatsu Ken,
Okano Hideyuki
Publication year - 2009
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2009.06256.x
Subject(s) - neural stem cell , embryonic stem cell , biology , lectin , neurosphere , microglia , microbiology and biotechnology , cell , neural cell , glycan , stem cell , glycoprotein , biochemistry , adult stem cell , immunology , gene , inflammation
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.