Rusenko K W, Stach R W
J Neurosci Res. 1982;8(2-3):263-70. doi: 10.1002/jnr.490080216.
Culture of embryonic chicken dorsal root ganglia for periods exceeding a week generally require serum supplementation and a trophic factor such as nerve growth factor. In this communication we describe the culture of chicken E-9 dorsal root ganglia for periods up to 2 weeks in a system that is composed only of a simple basic salts solution and 10 ng/ml (3.8 X 10(-10)M) beta nerve growth factor. Commercially available tissue culture dishes are used which have a hydrophilic, gas-diffusable membrane on the bottom to which the ganglia directly attach, eliminating the need for added substratum. Sparse fiber outgrowth appears after 24 hours and growth of the fibers continues for 120 hours of incubation. At this time, the fibers are extremely dense and reach approximately 3.5-4.0 diameters from the body of the ganglion. Continued survival of these fibers appears to depend on the concentration of nonneuronal cells present in the dish. No fiber outgrowth occurs at any time in the absence of beta nerve growth factor. The simplicity and purity of this culture system makes it an attractive tool in the study of primary cell-cell interactions, the production of trophic factors by nonneuronal cells, and biochemical and physiological analyses of growing neurons.
鸡胚背根神经节培养超过一周通常需要补充血清和诸如神经生长因子等营养因子。在本交流中,我们描述了在仅由简单的基础盐溶液和10 ng/ml(3.8×10⁻¹⁰M)β神经生长因子组成的系统中,将鸡E-9背根神经节培养长达2周的情况。使用市售的组织培养皿,其底部有一层亲水性、可气体扩散的膜,神经节可直接附着在上面,无需添加底物。24小时后出现稀疏的纤维生长,在孵育120小时期间纤维持续生长。此时,纤维极其密集,从神经节主体延伸约3.5 - 4.0个直径。这些纤维的持续存活似乎取决于培养皿中存在的非神经元细胞的浓度。在没有β神经生长因子的情况下,任何时候都不会出现纤维生长。这种培养系统的简单性和纯度使其成为研究原代细胞 - 细胞相互作用、非神经元细胞产生营养因子以及对生长中的神经元进行生化和生理分析的有吸引力的工具。