Neuroepithelial stem cells from the embryonic spinal cord: Isolation, characterization, and clonal analysis
Neuroepithelial stem cells from the embryonic spinal cord: Isolation, characterization, and clonal analysis
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DOI：
10.1006/dbio.1997.8592
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发表时间：
1997-06-15
影响因子：
2.7
通讯作者：
Rao, MS
中科院分区：
文献类型：
--
作者：
Kalyani, A;Hobson, K;Rao, MS
来自尾神经管的E10.5大鼠神经上皮细胞（NEP细胞）的贴壁培养物需要FGF（成纤维细胞生长因子）和CEE（鸡胚提取物）来增殖并在培养物中维持未分化的表型。表皮生长因子（EGF）不支持E10.5 NEP细胞在贴壁培养和NEP细胞不形成EGF依赖的神经球。然而，NEP细胞可以作为FGF依赖性神经球生长。NEP细胞表达巢蛋白，缺乏神经元和神经胶质亚系的所有谱系特异性标志物，在多次传代中保持其多能性，并且当在不存在CEE的情况下铺在层粘连蛋白上时可以分化成神经元、星形胶质细胞和少突胶质细胞。在克隆培养中，NEP细胞进行自我更新并产生大小从单个细胞到数千个细胞不等的集落。除少数单细胞克隆外，所有其他NEP衍生的克隆都含有一种以上的鉴定表型，超过40%的集落含有A2 B5、β-111微管蛋白和GFAP免疫反应性细胞。因此，NEP细胞是多能的，能够产生多种神经衍生物。NEP细胞也分化成运动神经元免疫反应胆碱乙酰转移酶（ChAT）和低亲和力神经营养因子受体（p75）在质量和克隆培养。双标记的克隆ChAT和神经胶质细胞，神经元，或少突胶质细胞谱系标记显示，运动神经元总是出现在与其他分化细胞的混合培养。因此，NEP细胞代表运动神经元和其他脊髓细胞的共同祖细胞。并对NEP细胞与其他神经干细胞的关系进行了讨论。(C)北京：科学出版社.
Adherent cultures of E10.5 rat neuroepithelial cells (NEP cells) from the caudal neural tube require FGF (fibroblast growth factor) and CEE (chick embryo extract) to proliferate and maintain an undifferentiated phenotype in culture. Epidermal growth factor (EGF) does not support E10.5 NEP cells in adherent culture and NEP cells do not form EGF-dependent neurospheres. NEP cells, however, can be grown as FGF-dependent neurospheres. NEP cells express nestin and lack all lineage-specific markers for neuronal and glial sublineages, retain their pleuripotent character over multiple passages, and can differentiate into neurons, astrocytes, and oligodendrocytes when plated on laminin in the absence of CEE. In clonal culture, NEP cells undergo self-renewal and generate colonies that vary in size from single cells to several thousand cells. With the exception of a few single-cell clones, all other NEP-derived clones contain more than one identified phenotype, with over 40% of the colonies containing A2B5, beta-111 tubulin, and GFAP-immunoreactive cells. Thus, NEP cells are multipotent and capable of generating multiple neural derivatives. NEP cells also differentiate into motoneurons immunoreactive for choline acetyl transferase (ChAT) and the low-affinity neurotrophin receptor (p75) in both mass and clonal culture. Double labeling of clones for ChAT and glial, neuronal, or oligodendrocytic lineage markers shows that motoneurons always arose in mixed cultures with other differentiated cells. Thus, NEP cells represent a common progenitor for motoneurons and other spinal cord cells. The relationship of NEP cells with other neural stem cells is discussed. (C) 1997 Academic Press.