Substrate requirement and media supplements necessary for the long-term survival of chick sympathetic and sensory neurons cultured without serum.
Substrate requirement and media supplements necessary for the long-term survival of chick sympathetic and sensory neurons cultured without serum.
复制标题
无血清培养的小鸡交感神经元和感觉神经元长期存活所需的底物要求和培养基补充。
DOI：
10.1016/0014-4827(82)90157-4
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发表时间：
1982
影响因子：
3.7
通讯作者：
Thoenen,H
中科院分区：
文献类型：
--
作者：
Wakade,AR;Edgar,D;Thoenen,H
从胚胎鸡交感神经和背根（感觉）神经节分离的神经元，从大多数的神经节非神经元细胞的preplating技术，然后培养无血清的聚鸟氨酸基板上。培养液为F14，添加5 μg/ml转铁蛋白、5 μg/ml胰岛素和10 ng/ml神经生长因子（NGF）。在这些条件下，神经元迅速粘附到基底上，从而避免了在血清存在下启动培养的需要。定量研究表明，在这种无血清培养基中存活的神经元数量与在含有NGF和血清的最佳条件下存活的神经元数量相同。在培养基中添加额外的补充剂，如黄体酮、腐胺、硒、三碘甲状腺原氨酸或羟考的松，并不能进一步提高神经元的存活率。然而，在本条件下的神经元的长期存活率比血清存在下的神经元的长期存活率提高。此外，少数非神经元细胞保留在预铺板后的文化被发现在长期的培养期间广泛增殖。
Neurons dissociated from embryonic chick sympathetic and dorsal root (sensory) ganglia were separated from the majority of the ganglionic non-neuronal cells by a preplating technique and then cultured without serum on a polyornithine substrate. The culture medium was F14, supplemented with 5 μg/ml transferrin, 5 μg/ml insulin and 10 ng/ml nerve growth factor (NGF). Under these conditions the neurons rapidly adhered to the substrate, thus obviating the need to initiate the culture in the presence of serum. Quantitative studies showed that the numbers of neurons surviving in this serum-free medium were the same as those surviving under optimal conditions with NGF and serum. Additional supplements to the culture medium—such as progesterone, putrescine, selenium, triiodothyronine, or hydroxycortisone—did not enhance neuronal survival further. The long-term survival of the neurons under the present conditions was, however, improved over that in the presence of serum. Additionally, the few non-neuronal cells remaining in the cultures after preplating were found to proliferate extensively during long culture periods.