Center for Vitamins and Cancer Research, Department of Radiology, School of Medicine, University of Colorado, Health Sciences Center, 4200 E. 9th Avenue, Denver, CO 80262-0276, USA.
Restor Neurol Neurosci. 1994 Jan 1;7(1):13-9. doi: 10.3233/RNN-1994-7103.
The use of fetal central nervous system (CNS) tissue in neural transplants has ethical, availability and some biological limitations. In order to overcome these problems, homogeneous populations of specific neurons in vitro should be established. Transformed nerve cells such as neuroblastoma cells (NBP2) in culture can serve as one of the sources of donor neurons in neural transplants provided they can be induced to differentiate terminally. This study showed that treatment of murine neuroblastoma (NBP2) cells with 4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone (R020-1724, an inhibitor of cyclic nucleotide phosphodiesterase), and β-carotene for a period of 3 days followed by X-irradiation with 20 Gy or above produced 100% terminally differentiated cells. These differentiated NB cells had long and extensive neurites, contained elevated levels of tyrosine hydroxylase (TH) activity and very low levels of MHC class I and II antigens, and were non-tumorigenic. The viability of differentiated NB cells when determined on the criteria of attachment efficiency, re-extension of neurites and presence of TH after replating was only 9%. This was in contrast to the trypan blue exclusion technique which showed that over 90% of differentiated NB cells in culture were viable.
在神经移植中使用胎儿中枢神经系统 (CNS) 组织存在伦理、可用性和一些生物学限制。为了克服这些问题，应该在体外建立特定神经元的同质群体。转化的神经细胞，如神经母细胞瘤细胞 (NBP2)，在培养中可以作为神经移植中供体细胞的来源之一，前提是它们可以被诱导终末分化。本研究表明，用 4-(3-正丁氧基-4-甲氧基苄基)-2-咪唑烷酮（R020-1724，环核苷酸磷酸二酯酶抑制剂）和β-胡萝卜素处理小鼠神经母细胞瘤（NBP2）细胞 3 天，然后用 20 Gy 或更高剂量的 X 射线照射，可产生 100%的终末分化细胞。这些分化的 NB 细胞具有长而广泛的轴突，酪氨酸羟化酶 (TH) 活性升高，MHC Ⅰ类和Ⅱ类抗原水平极低，且无致瘤性。根据附着效率、神经突再延伸和再接种后存在 TH 的标准确定的分化 NB 细胞的存活率仅为 9%。这与台盼蓝排除技术形成对比，后者显示培养中的分化 NB 细胞中超过 90%是存活的。
