Cell surface N-glycans mediated isolation of mouse neural stem cells

Makoto Hamanoue, Yumi Matsuzaki, Ken Ichiro Sato, Hirotaka James Okano, Shinsuke Shibata, Isamu Sato, Sadafumi Suzuki, Miyuki Ogawara, Ken Takamatsu, Hideyuki Okano

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)1575-1584
Number of pages10
JournalJournal of Neurochemistry
Volume110
Issue number5
DOIs
Publication statusPublished - 2009 Sep

Fingerprint

Neural Stem Cells
Stem cells
Polysaccharides
Brain
Lectins
Asparagine
Surface Antigens
Oligosaccharides
Labeling
Neurons
Deterioration
Glycoproteins
Oligodendroglia
Microglia
Phytohemagglutinins
Membranes
erythroagglutinating phytohemagglutinin
Phenotype

Keywords

  • Cell separation
  • Flow cytometry
  • Membrane glycoproteins
  • Phytohemagglutinins
  • Plant lectins
  • Stem cells

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Cell surface N-glycans mediated isolation of mouse neural stem cells. / Hamanoue, Makoto; Matsuzaki, Yumi; Sato, Ken Ichiro; Okano, Hirotaka James; Shibata, Shinsuke; Sato, Isamu; Suzuki, Sadafumi; Ogawara, Miyuki; Takamatsu, Ken; Okano, Hideyuki.

In: Journal of Neurochemistry, Vol. 110, No. 5, 09.2009, p. 1575-1584.

Research output: Contribution to journalArticle

Hamanoue, M, Matsuzaki, Y, Sato, KI, Okano, HJ, Shibata, S, Sato, I, Suzuki, S, Ogawara, M, Takamatsu, K & Okano, H 2009, 'Cell surface N-glycans mediated isolation of mouse neural stem cells', Journal of Neurochemistry, vol. 110, no. 5, pp. 1575-1584. https://doi.org/10.1111/j.1471-4159.2009.06256.x
Hamanoue, Makoto ; Matsuzaki, Yumi ; Sato, Ken Ichiro ; Okano, Hirotaka James ; Shibata, Shinsuke ; Sato, Isamu ; Suzuki, Sadafumi ; Ogawara, Miyuki ; Takamatsu, Ken ; Okano, Hideyuki. / Cell surface N-glycans mediated isolation of mouse neural stem cells. In: Journal of Neurochemistry. 2009 ; Vol. 110, No. 5. pp. 1575-1584.
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