Production of compartmented cultures of rat sympathetic neurons
Production of compartmented cultures of rat sympathetic neurons
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DOI：
10.1038/nprot.2009.210
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发表时间：
2009-01-01
期刊：
影响因子：
14.8
通讯作者：
Mok, Sue-Ann
中科院分区：
文献类型：
--
作者：
Campenot, Robert B.;Lund, Karen;Mok, Sue-Ann
隔室培养，其中在近端隔室中铺板的原代神经元将其轴突发送到硅脂屏障下并进入左右远端隔室，增强了神经元培养物的实验能力。治疗可以分别应用于细胞体/近端轴突或远端轴突，并且细胞体/近端轴突和远端轴突可以分别收获和分析。远端轴突可以被切断，并且神经元可以在其轴突再生的同时被研究。培养皿的构建需要3 h进行48次培养，神经元的制备也需要3 h。隔室培养物为生物化学分析（如免疫印迹）提供了足够的细胞材料。隔室培养物的用途包括神经营养因子逆行信号传导、轴突运输、轴突蛋白和脂质生物合成的研究。在这里，我们专注于从新生大鼠培养的交感神经元，并提供协议的生产和一些分区文化的用途。
The compartmented culture, in which primary neurons plated in a proximal compartment send their axons under silicone grease barriers and into left and right distal compartments, has enhanced the experimental capabilities of neuronal cultures. Treatments can be applied separately to cell bodies/proximal axons or distal axons, and cell bodies/proximal axons and distal axons can be separately harvested and analyzed. Distal axons can be axotomized, and the neurons can be studied while their axons regenerate. Construction of the culture dishes requires 3 h for 48 cultures, and preparing the neurons also requires 3 h. Compartmented cultures provide enough cellular material for biochemical analyses such as immunoblotting. The uses of compartmented cultures have included studies of neurotrophic factor retrograde signaling, axonal transport, and axonal protein and lipid biosynthesis. Here we focus on sympathetic neurons cultured from neonatal rats and provide protocols for the production and some of the uses of compartmented cultures.