Sensenbrenner M, Maderspach K, Latzkovits L, Jaros G G
Dev Neurosci. 1978;1(2):90-101. doi: 10.1159/000112560.
Dissociated cells from 5- to 12-day-old chick embryo cerebral hemispheres were cultivated in polylysine-coated plastic Petri dishes. The polylysine substrate was observed to be favorable for the growth of neuronal cells, whereas glioblast proliferation was inhibited. The optimal conditions for the production of a predominantly neuronal culture were to use cerebral hemispheres from 7-day-old chick embryos, to dissociate the brain tissue mechanically and to seed the cells at a concentration range between 1.5 and 5 X 10(6) cells/ml. The cultures were observed by phase contrast microscopy. Most cells grew fibers and differentiated into bipolar and multipolar neurons. These neurons were stained by thionine, which demonstrated the presence of Nissl bodies. The silver impregnation revealed the presence of neurofibrils within the nerve fibers. Acetylcholinesterase was found to be present in the neuronal cells, but absent in the glioblasts. Under our culture conditions the neurons survived for 10-12 days. This system should allow further studies on the effects of growth factors on the differentiation of isolated neurons as well as investigations on neuron-glial interrelationship.
将5至12日龄鸡胚大脑半球的解离细胞培养于聚赖氨酸包被的塑料培养皿中。观察到聚赖氨酸底物有利于神经元细胞的生长，而成胶质细胞的增殖受到抑制。产生以神经元为主的培养物的最佳条件是使用7日龄鸡胚的大脑半球，机械解离脑组织，并将细胞以1.5至5×10(6)个细胞/毫升的浓度接种。通过相差显微镜观察培养物。大多数细胞长出纤维并分化为双极和多极神经元。这些神经元用硫堇染色，显示有尼氏体存在。银浸染显示神经纤维内有神经原纤维。发现乙酰胆碱酯酶存在于神经元细胞中，但在成胶质细胞中不存在。在我们的培养条件下，神经元存活10至12天。该系统应能进一步研究生长因子对分离神经元分化的影响以及对神经元-神经胶质细胞相互关系的研究。
