PRIMARY CULTURES OF DISSOCIATED SYMPATHETIC NEURONS .1. ESTABLISHMENT OF LONG-TERM GROWTH IN CULTURE AND STUDIES OF DIFFERENTIATED PROPERTIES
PRIMARY CULTURES OF DISSOCIATED SYMPATHETIC NEURONS .1. ESTABLISHMENT OF LONG-TERM GROWTH IN CULTURE AND STUDIES OF DIFFERENTIATED PROPERTIES
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DOI：
10.1083/jcb.59.2.329
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发表时间：
1973-01-01
影响因子：
7.8
通讯作者：
PATTERSON, PH
中科院分区：
文献类型：
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作者：
MAINS, RE;PATTERSON, PH
机械搅动破坏大鼠交感神经节产生游离的原代神经元，并观察分离神经元长期培养的条件。通过在培养过程中添加或删除碳酸氢盐，神经元可以与非神经细胞一起或不一起生长。荧光组化显示离体神经元含有儿茶酚胺；用放射性前体孵育来验证多巴胺和去甲肾上腺素的合成和积累。神经元也以酪胺为前体产生章鱼胺，但不以酪氨酸为前体。在叶丝氨酸存在的情况下，古老的培养物合成并储存了少量的乙酰胆碱。培养物不合成和积累可检测水平的放射性y-氨基丁酸、5-羟色胺或组胺。组织培养系统，由长期培养的分离的原代神经元组成，具有已知的同质起源和功能，可以在存在或缺乏其他细胞类型的情况下生长。有几种培养系统可以使用来自不同胚胎来源的神经元来满足这些标准（背根神经节：Scott et al., 1969, Miller et al., 1970, Okun, 1972；大脑：Varon and Raiborn, 1969；脊髓：Fischbach， 1972）。这些神经元可以通过使用分离程序或抗有丝分裂剂在几乎没有非神经细胞的情况下生长。也有明显来源于神经的肿瘤细胞（Augusti-Tocco and Sato, 1969; Schubert et al., 1969）。
Rat sympathetic ganglia were disrupted by mechanical agitation to yield dissociated primary neurons, and the conditions for long-term growth in culture of the isolated neurons were examined. The neurons were grown with or without non-neural cells, simply by the addition or deletion of bicarbonate during growth in culture. Fluorescence histochemistry indicated that the isolated neurons contained catecholamines; incubations with radioactive precursors were used to verify the synthesis and accumulation of both dopamine and norepinephrine. The neurons also produced octopamine using tyramine as precursor, but not with tyrosine as the precursor. In the presence of eserine, older cultures synthesized and stored small amounts of acetylcholine. The cultures did not synthesize and accumulate detectable levels of radioactive y-aminobutyric acid, 5-hydroxytryptamine, or histamine. tissue culture system consisting of long-term cultures of dissociated primary neurons of known homogeneous origin and function which could be grown in the presence or absence of other cell types. There are several culture systems which meet some of these criteria using neurons from various embryonic sources (dorsal root ganglia: Scott et al., 1969, Miller et al., 1970, Okun, 1972; cerebrum: Varon and Raiborn, 1969; spinal cord: Fischbach, 1972). These neurons can be grown in the virtual absence of non-neural cells by use of separation procedures or antimitotic agents. Also available are tumor cells apparently of nervous origin (Augusti-Tocco and Sato, 1969; Schubert et al., 1969).