Cell surface N-glycans mediated isolation of mouse neural 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.
Hamanoue
Makoto
M
Department of Physiology, Toho University School of Medicine, 5-21-16 Ohmori-nishi, Ohta-ku, Tokyo 143-8540, Japan. hamanoue-ns@umin.ac.jp
Matsuzaki
Yumi
Y
Sato
Ken-Ichiro
K
Okano
Hirotaka James
HJ
Shibata
Shinsuke
S
Sato
Isamu
I
Suzuki
Sadafumi
S
Ogawara
Miyuki
M
Takamatsu
Ken
K
Okano
Hideyuki
H
D000818:Animals; D002462:Cell Membrane / Q000502:physiology*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology*; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D051379:Mice; D008813:Mice, Inbred ICR; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D011134:Polysaccharides / Q000502:physiology*; D011247:Pregnancy
D011134:Polysaccharides