Tuttle J B, Suszkiw J B, Ard M
Brain Res. 1980 Feb 3;183(1):161-80. doi: 10.1016/0006-8993(80)90127-4.
Ciliary ganglion neurons from chick embryo were grown in cell culture following enzymatic and mechanical dissociation. The culture conditions necessary for long-term (greater than 21 days) survival of the isolated neurons were investigated. Neurons could be cultured with and without non-neural cells by adjusting the culture substrate. Chick embryo extract was found to be essential in the growth medium, and the inclusion of horse serum had an additional beneficial effect. Also, non-neural cells increased the survival of these neurons in culture. Thus, there appear to be several factors involved in the survival of these neurons in culture. Assay of choline acetyltransferase activity revealed a 100-fold increase in activity over the first two weeks in vitro, and the developmental pattern of the enzyme activity was initially similar to that seen in vivo. The cultured neurons also retained many of the electrophysiological properties of ganglionic neurons in vivo. These included normal resting transmembrane potential, action potential amplitude and an afterpotential. The passive membrane properties of the cultured neurons (membrane resistance, capacitance and time constant) differed somewhat from those of neurons in ganglia. These results suggest that if proper culture conditions are provided, these parasympathetic neurons adapt well to cell culture and develop many of the properties of normal ciliary ganglion neurons in vivo.
经酶解和机械解离后，将鸡胚睫状神经节神经元进行细胞培养。研究了分离神经元长期（超过21天）存活所需的培养条件。通过调整培养底物，神经元可以与非神经细胞一起或不与非神经细胞一起培养。发现鸡胚提取物在生长培养基中必不可少，添加马血清有额外的有益作用。此外，非神经细胞可提高这些神经元在培养中的存活率。因此，这些神经元在培养中的存活似乎涉及多个因素。胆碱乙酰转移酶活性测定显示，在体外的头两周内活性增加了100倍，并且该酶活性的发育模式最初与体内所见相似。培养的神经元还保留了体内神经节神经元的许多电生理特性。这些特性包括正常的静息跨膜电位、动作电位幅度和后电位。培养神经元（膜电阻、电容和时间常数）的被动膜特性与神经节中的神经元略有不同。这些结果表明，如果提供适当的培养条件，这些副交感神经元能很好地适应细胞培养，并发展出体内正常睫状神经节神经元的许多特性。
