Monnet-Tschudi F, Honegger P
Institut de Physiologie, Faculté de Médecine, Université de Lausanne, Switzerland.
Dev Neurosci. 1989;11(1):30-40. doi: 10.1159/000111883.
Maturation of astrocytes, neurons, and oligodendrocytes was studied in serum-free aggregating cell cultures of fetal rat telencephalon by an immunocytochemical approach. Cell type-specific immunofluorescence staining was examined by using antibodies directed against glial fibrillary acidic protein (GFAP) and vimentin, two astroglial markers; neuron-specific enolase (NSE) and neurofilament (NF), two neuronal markers, and galactocerebroside (GC), an oligodendroglial marker. It was found that the cellular maturation in aggregates is characterized by distinct developmental increases in immunoreactivity for GFAP, vimentin, NSE, NF, and GC, and by a subsequent decrease of vimentin-positive structures in more differentiated cultures. These findings are in agreement with observations in vivo, and they corroborate previous biochemical studies of this histotypic culture system. Treatment of very immature cultures with a low dose of epidermal growth factor (EGF, 5 ng/ml) enhanced the developmental increase in GFAP, NSE, NF and GC immunoreactivity, suggesting an acceleration of neuronal and glial maturation. In addition, EGF was found to alter the cellular organization within the aggregates, presumably by influencing cell migration.
通过免疫细胞化学方法,在胎鼠端脑的无血清聚集细胞培养物中研究了星形胶质细胞、神经元和少突胶质细胞的成熟情况。使用针对胶质纤维酸性蛋白(GFAP)和波形蛋白这两种星形胶质细胞标志物、神经元特异性烯醇化酶(NSE)和神经丝(NF)这两种神经元标志物以及半乳糖脑苷脂(GC)这种少突胶质细胞标志物的抗体,检测细胞类型特异性免疫荧光染色。结果发现,聚集体中的细胞成熟表现为GFAP、波形蛋白、NSE、NF和GC免疫反应性明显的发育性增加,以及在更分化的培养物中波形蛋白阳性结构随后减少。这些发现与体内观察结果一致,并证实了此前对这种组织型培养系统的生化研究。用低剂量表皮生长因子(EGF,5 ng/ml)处理非常不成熟的培养物,可增强GFAP、NSE、NF和GC免疫反应性的发育性增加,表明神经元和胶质细胞成熟加速。此外,发现EGF可改变聚集体内的细胞组织,可能是通过影响细胞迁移实现的。