Properties of a fetal multipotent neural stem cell (NEP cell)
Properties of a fetal multipotent neural stem cell (NEP cell)
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DOI：
10.1006/dbio.2002.0828
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发表时间：
2002-11-15
影响因子：
2.7
通讯作者：
Rao, MS
中科院分区：
文献类型：
--
作者：
Cai, JL;Wu, YY;Rao, MS
对发育中的神经管中存在的多潜能神经干细胞(NSCs)(E10.5，神经上皮细胞；NEP)进行了候选干细胞标志物的表达检测，并将这些标志物的表达与后来出现的前体细胞(E14.5)进行了比较，后者可以通过胚胎神经细胞黏附分子(E-NCAM)和A2B5的表达来区分。NEP细胞有缝隙连接，表达连接蛋白，似乎缺乏长纤毛。大多数候选标记，包括巢蛋白、早老素、Notch和Numb，在NEP细胞和其他细胞群体中都有表达。免疫细胞化学和原位杂交显示成纤维细胞生长因子受体4(FGFR4)、Frizzled9(Fz9)和SRY盒基因2(Sox2)是神经干细胞区别于其他前体细胞的标志物。Hoechst123和罗丹明33342染色、端粒酶表达、端粒酶活性或断裂点簇区蛋白I(Bcrp1)的表达均不能区分NEP干细胞和其他分裂细胞。然而，NEP细胞缺乏几个谱系标记的表达，这些标记是由后来出现的细胞表达的。这些包括CD44、E-NCAM、A2B5、表皮生长因子受体(EGFR)和血小板衍生生长因子受体-α(PDGFRpha)的表达缺失，这表明使用细胞表面表位的负选择可以用于从混合细胞培养中分离干细胞群体。使用从E14.5期胚胎分离的细胞的混合培养，我们证明可以通过耗尽表达E-NCAM或A2B5免疫反应性的分化细胞来丰富NEP细胞。总体而言，我们的结果表明，结合使用的一系列标志物可以可靠地将多潜能神经干细胞与其他前体细胞以及中枢神经系统中存在的分化细胞区分开来。(C)2002年爱思唯尔科学公司(美国)
Multipotent neural stem cells (NSCs) present in the developing neural tube (E10.5, neuroepithelial cells; NEP) were examined for the expression of candidate stem cell markers, and the expression of these markers was compared with later appearing precursor cells (E14.5) that can be distinguished by the expression of embryonic neural cell adhesion molecule (E-NCAM) and A2B5. NEP cells possess gap junctions, express connexins, and appear to lack long cilia. Most candidate markers, including Nestin, Presenilin, Notch, and Numb, were expressed by both NEP cells as well as other cell populations. Fibroblast growth factor receptor 4 (FGFR4), Frizzled 9 (Fz9), and SRY box-containing gene 2 (Sox2) as assessed by immunocytochemistry and in situ hybridization are markers that appear to distinguish NSCs from other precursor cells. Neither Hoechst 33342 nor rhodamine- 123 staining, telomerase (Tert) expression, telomerase activity, or breakpoint cluster region protein I (Bcrp1) transporter expression could be used to distinguish NEP stem cells from other dividing cells. NEP cells, however, lacked expression of several lineage markers that are expressed by later appearing cells. These included absence of expression of CD44, E-NCAM, A2B5, epidermal growth factor receptor (EGFR), and platelet-derived growth factor receptor-alpha (PDGFRalpha), suggesting that negative selection using cell surface epitopes could be used to isolate stem cell populations from mixed cultures of cells. Using mixed cultures of cells isolated from E14.5 stage embryos, we show that NEP cells can be enriched by depleting differentiating cells that express E-NCAM or A2B5 immunoreactivity. Overall, our results show that a spectrum of markers used in combination can reliably distinguish multipotent NSCs from other precursor cells as well as differentiated cells present in the CNS. (C) 2002 Elsevier Science (USA)