Mørk S J, May E E, Papasozomenos S C, Vinores S A
Neuropathol Appl Neurobiol. 1986 May-Jun;12(3):277-89. doi: 10.1111/j.1365-2990.1986.tb00140.x.
The human medulloblastoma cell line, TE-671, was studied in vitro both in monolayer culture and in a three-dimensional culture system using gelfoam as the supporting matrix. Flow cytometry studies of cells grown in monolayer culture revealed a unimodal, tetraploid DNA content. Most cells in both in vitro systems contained neuron-specific enolase (NSE), actin, and tubulin, while only occasional cells or cell clusters contained the 68,000 molecular weight subunit of neurofilaments (NF mol. wt 68,000) or microtubule-associated protein 2 (MAP-2). In monolayer culture, long cellular processes containing NSE, NF mol. wt 68,000 and MAP-2, which were present at 2 days, were nearly absent by 7 days. All antigens were present at 4 days in the organ culture system; by 72 days, cells still stained positively for NF mol. wt 68 000 and MAP-2, but staining for NSE, actin, and beta-tubulin was diminished as compared to 4 days. Retinoic acid (RA) in the 13-cis isomer form at 10(-6) M was applied to monolayer cultures at day 1 for 6 days and to gelfoam cultures at day 1 for 28 days. RA did not significantly alter cell proliferation up to 7 days in vitro and did not appreciably affect cellular expression of NSE, NF mol. wt 68 000, MAP-2, beta-tubulin, or actin in either system. By electron microscopy, most cells grown under different culture conditions with or without RA treatment appeared to be undifferentiated and polygonal, with occasional cytoplasmic annulate lamellae. The immunohistochemical and ultrastructural features reported indicate that the TE-671 medulloblastoma line is composed primarily of primitive neuroepithelial cells with a limited potential for neuronal differentiation. This differentiation was not promoted by RA or by an in vitro system known to favour differentiation in a number of human and animal nervous system tumours. The findings suggest that the cells of the TE-671 line lack either receptors for retinoic acid or the capacity to respond to bound retinoic acid.
对人髓母细胞瘤细胞系TE - 671进行了体外研究,分别采用单层培养和以明胶海绵作为支持基质的三维培养系统。对单层培养的细胞进行流式细胞术研究显示其DNA含量呈单峰四倍体。在这两种体外培养系统中,大多数细胞都含有神经元特异性烯醇化酶(NSE)、肌动蛋白和微管蛋白,而只有偶尔的细胞或细胞簇含有分子量为68,000的神经丝亚基(NF分子量68,000)或微管相关蛋白2(MAP - 2)。在单层培养中,含有NSE、NF分子量68,000和MAP - 2的长细胞突起在第2天出现,到第7天几乎消失。在器官培养系统中,所有抗原在第4天均存在;到第72天,细胞对NF分子量68,000和MAP - 2仍呈阳性染色,但与第4天相比,NSE、肌动蛋白和β - 微管蛋白的染色减弱。在第1天将10(-6) M的13 - 顺式视黄酸(RA)应用于单层培养6天,应用于明胶海绵培养28天。在体外培养至7天,RA并未显著改变细胞增殖,且在任一系统中均未明显影响NSE、NF分子量68,000、MAP - 2、β - 微管蛋白或肌动蛋白的细胞表达。通过电子显微镜观察,在不同培养条件下生长且经或未经RA处理的大多数细胞似乎未分化且呈多边形,偶尔可见胞质环状片层。所报道的免疫组织化学和超微结构特征表明,TE - 671髓母细胞瘤细胞系主要由具有有限神经元分化潜能的原始神经上皮细胞组成。RA或已知有利于多种人类和动物神经系统肿瘤分化的体外系统均未促进这种分化。这些发现表明,TE - 671细胞系的细胞要么缺乏视黄酸受体,要么缺乏对结合的视黄酸作出反应的能力。