Galiana E, Bernard R, Borde I, Rouget P, Evrard C
Laboratoire Biologie Moléculaire et Différenciation, Université Paris-6 et Collège de France.
J Neurosci Res. 1993 Oct 1;36(2):133-46. doi: 10.1002/jnr.490360204.
Bipotent glial progenitors have been immortalized by the transfer of the adenovirus E1A gene into primary cultured cells from embryonic rat brain. The lines obtained are phenotypically untransformed, retain growth contact-inhibition, and are able to differentiate, unless they are surtransfected with transforming oncogenes. Depending on the growth conditions, these immortalized cells express differentially either oligodendrocyte or astrocyte-specific markers and genes. After being seeded in serum-free medium, they display gangliosides recognized by A2B5 monoclonal antibody, and then they express sequentially O4 epitopes, galactocerebroside, and the myelin protein DM20. When grown in serum-supplemented medium, the cells express at first A2B5 epitopes, and then transiently O4 and galactocerebroside; after reaching confluence, O4 and galactocerebroside become undetectable, whereas the cells begin to coexpress glial fibrillary acidic protein and glutamine synthetase. These results indicate that the cell lines can undergo a differentiation reminiscent both of O-2A progenitors and of plastic process-bearing glial subpopulations. The cells were also genetically marked by the stable introduction of the nlslacZ reporter gene. Thus, the lines could be useful for studying direct interactions in vitro, or for post-grafting investigations. They should also provide a model for studying the mechanisms involved in the commitment and in the control of proliferation and differentiation of this cell lineage. This suggestion is consistent with the data indicating a growth arrest-dependent differential expression of a novel gene encoding a protein with a helix-loop-helix domain.
通过将腺病毒E1A基因导入来自胚胎大鼠脑的原代培养细胞，双能神经胶质祖细胞已被永生化。所获得的细胞系在表型上未发生转化，保留生长接触抑制，并且能够分化，除非它们被转化癌基因超转染。根据生长条件，这些永生化细胞差异表达少突胶质细胞或星形胶质细胞特异性标志物和基因。接种于无血清培养基后，它们表达被A2B5单克隆抗体识别的神经节苷脂，然后依次表达O4表位、半乳糖脑苷脂和髓磷脂蛋白DM20。当在补充血清的培养基中生长时，细胞首先表达A2B5表位，然后短暂表达O4和半乳糖脑苷脂；汇合后，O4和半乳糖脑苷脂变得不可检测，而细胞开始共表达胶质纤维酸性蛋白和谷氨酰胺合成酶。这些结果表明，这些细胞系能够经历一种既类似于少突胶质前体细胞又类似于具有可塑性突起的神经胶质亚群的分化过程。通过稳定导入nlslacZ报告基因对细胞进行了基因标记。因此，这些细胞系可用于研究体外直接相互作用或移植后研究。它们还应为研究该细胞谱系的定向分化以及增殖和分化控制机制提供一个模型。这一推测与数据一致，这些数据表明一种编码具有螺旋-环-螺旋结构域蛋白质的新基因的表达存在生长停滞依赖性差异。
