Kuhn Thomas B
Department of Pharmaceutical Sciences, University of Montana, Missoula, Montana 59812, USA.
Methods Cell Biol. 2003;71:67-87. doi: 10.1016/s0091-679x(03)01005-7.
The chick embryo has a long tradition as a model organism in developmental biology as well as embryology. A year-round supply of fertilized eggs, accessibility to all stages of development, and the ease of manipulation of the embryo all contribute to the advantages of investigations using chick embryos. A plethora of culture systems have been developed over the past century allowing to culture intact embryos from as early as 2 days of development. Other culture systems include whole embryo slices, organotypic cultures, tissue explants, and dissociated cultures. Studies utilizing the chick embryo, and in particular spinal motor neurons, were crucial for our present knowledge of the development but also adult physiology, injury, and disease of the nervous system. Extensive studies on spinal motor neurons revealed many molecular mechanisms underlying fundamental events, such as neural induction, axon guidance, programmed cell death, and neuron-target interaction. Cultures of dissociated spinal motor neurons represent one important experimental paradigm. This chapter describes two alternative procedures to establish dissociated spinal motor neuron cultures with virtually no contamination by nonneuronal cells.
鸡胚作为发育生物学和胚胎学中的一种模式生物有着悠久的传统。全年供应的受精卵、可接触到发育的各个阶段以及易于操作胚胎,这些都构成了使用鸡胚进行研究的优势。在过去的一个世纪里,已经开发出了大量的培养系统,能够从发育早期的第2天开始培养完整的胚胎。其他培养系统包括全胚胎切片、器官型培养、组织外植体和离散培养。利用鸡胚,特别是脊髓运动神经元进行的研究,对于我们目前关于神经系统发育、成年生理学、损伤和疾病的知识至关重要。对脊髓运动神经元的广泛研究揭示了许多基本事件背后的分子机制,如神经诱导、轴突导向、程序性细胞死亡和神经元与靶标的相互作用。离散的脊髓运动神经元培养是一种重要的实验范式。本章描述了两种建立离散脊髓运动神经元培养物的替代方法,几乎没有非神经元细胞的污染。