Kalyani A, Hobson K, Rao M S
Department of Neurobiology and Anatomy, University of Utah School of Medicine, Salt Lake City 84132, USA.
Dev Biol. 1997 Jun 15;186(2):202-23. doi: 10.1006/dbio.1997.8592.
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.
来自尾神经管的E10.5大鼠神经上皮细胞（NEP细胞）的贴壁培养物需要成纤维细胞生长因子（FGF）和鸡胚提取物（CEE）才能在培养中增殖并维持未分化表型。表皮生长因子（EGF）不支持E10.5 NEP细胞的贴壁培养，且NEP细胞不能形成依赖EGF的神经球。然而，NEP细胞可以作为依赖FGF的神经球生长。NEP细胞表达巢蛋白，缺乏神经元和神经胶质亚谱系的所有谱系特异性标志物，在多次传代后仍保持其多能特性，并且在无CEE的情况下接种在层粘连蛋白上时可分化为神经元、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形成大小从单细胞到数千个细胞不等的集落。除了少数单细胞克隆外，所有其他源自NEP的克隆都包含不止一种已鉴定的表型，超过40%的集落含有A2B5、β-III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。在批量培养和克隆培养中，NEP细胞还能分化为对胆碱乙酰转移酶（ChAT）和低亲和力神经营养因子受体（p75）呈免疫反应性的运动神经元。对ChAT与神经胶质、神经元或少突胶质细胞谱系标志物的克隆进行双重标记显示，运动神经元总是在与其他分化细胞的混合培养物中出现。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。本文讨论了NEP细胞与其他神经干细胞的关系。
