Kim Hyun Jeong, Magrané Jordi
Department of Neurology and Neuroscience, Weill Cornell Medical College, New York, NY, USA.
Methods Mol Biol. 2011;793:63-75. doi: 10.1007/978-1-61779-328-8_4.
Many experimental animal models of human neurodegenerative diseases have been developed to understand the events leading toward neuronal dysfunction and death. However, definitive comprehension of the molecular and cellular mechanisms in these animal models is problematic because of the complexity of the intact nervous tissue. Primary neuronal cultures prepared from rodent nervous tissues represent a powerful tool not only to study the individual contribution of different cell types (such as neurons or glia) to disease progression, but also to investigate the role of neuron-glia interactions during development and pathogenesis of disease. Here, we describe a method to isolate and culture neurons and astrocytes from the mouse cerebral cortex, and we also present a practical application for transfection and subsequent immunofluorescence.
为了了解导致神经元功能障碍和死亡的相关事件,人们已经建立了许多人类神经退行性疾病的实验动物模型。然而,由于完整神经组织的复杂性,对这些动物模型中分子和细胞机制的明确理解存在问题。从啮齿动物神经组织制备的原代神经元培养物是一种强大的工具,不仅可以研究不同细胞类型(如神经元或神经胶质细胞)对疾病进展的个体贡献,还可以研究神经元-神经胶质细胞相互作用在疾病发生发展过程中的作用。在此,我们描述了一种从小鼠大脑皮层分离和培养神经元及星形胶质细胞的方法,并且还展示了转染及后续免疫荧光的实际应用。