Vicario-Abejón Carlos
Centro de Investigaciones Biológicas, Censejo Superior de Investigaciones Cientificas, Madrid, Spain.
Curr Protoc Neurosci. 2004 May;Chapter 3:Unit 3.2. doi: 10.1002/0471142301.ns0302s26.
In culture, hippocampal cells can develop to express neuronal antigens and acquire mature neuronal morphologies, including axons, complex dendritic trees, and synapses that are electrophysiologically active. This system is suitable for studying neuronal differentiation and other events, such as synaptogenesis. It is also a valuable model for investigating synaptic plasticity and exploring the mechanisms of neuronal degeneration. This unit provides a protocol for culturing neurons prepared from embryonic (E-18) rat or mouse hippocampus, but could also be used to grow neurons from embryonic cortex, olfactory bulb, striatum, or spinal cord. A second method is included for preparing neuronal cultures from embryos with different genotypes, such as those from transgenic mice. Also described is the preparation of polyornithine- and fibronectin-coated coverslips, which are highly adhesive and promote neurite outgrowth, for use in the culture protocols.
在培养过程中,海马体细胞能够发育并表达神经元抗原,形成成熟的神经元形态,包括轴突、复杂的树突分支以及具有电生理活性的突触。该系统适用于研究神经元分化及其他事件,如突触形成。它也是研究突触可塑性和探索神经元变性机制的宝贵模型。本单元提供了一种培养源自胚胎(E-18)大鼠或小鼠海马体的神经元的方法,也可用于培养源自胚胎皮质、嗅球、纹状体或脊髓的神经元。还介绍了第二种方法,用于从具有不同基因型的胚胎(如转基因小鼠胚胎)制备神经元培养物。此外,还描述了聚鸟氨酸和纤连蛋白包被盖玻片的制备方法,这些盖玻片具有高粘附性并能促进神经突生长,可用于培养方案。