Cell surface N-glycans mediated isolation of mouse neural stem cells
Cell surface N-glycans mediated isolation of mouse neural stem cells
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DOI：
10.1111/j.1471-4159.2009.06256.x
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发表时间：
2009-09-01
影响因子：
4.7
通讯作者：
Okano, Hideyuki
中科院分区：
文献类型：
--
作者：
Hamanoue, Makoto;Matsuzaki, Yumi;Okano, Hideyuki
由于缺乏有效的细胞表面标记，以及长期体外培养可能导致表型恶化，阻碍了从脑中分离神经干细胞(NSCs)。然而，在神经干细胞上几乎没有合适的特定细胞表面抗原可用于其未来的分离。用菜豆凝集素(E-PHA)与二天线复合型N-糖链结合，可检测到复合型天冬酰胺连接低聚糖(N-GAN)在胚胎和成人脑组织分离的脑细胞上的表达，E-PHA优先与纯化的NSCs结合，而不与神经元、小胶质细胞或少突胶质前体细胞结合。用E-PHA标记分离的小鼠胚胎脑细胞或成年脑细胞，可以使神经干细胞的神经球形成细胞的数量分别是未分离细胞的25倍或9倍。此外，凝集素印迹分析显示，在增殖的神经干细胞的膜部分中存在E-PHA识别的几种糖蛋白，但在分化的细胞中不存在。这些结果表明，复合型N-葡聚糖对于胚胎和成体脑中活的小鼠神经干细胞来说是一个有价值的细胞表面标志。
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.