Molecular and Cellular Neurobiotechnology, Institute for Bioengineering of Catalonia, Science Park of Barcelona, Barcelona, Spain.
Nat Protoc. 2012 Jan 19;7(2):268-80. doi: 10.1038/nprot.2011.445.
This protocol uses rat tail-derived type I collagen hydrogels to analyze key processes in developmental neurobiology, such as chemorepulsion and chemoattraction. The method is based on culturing small pieces of brain tissue from embryonic or early perinatal mice inside a 3D hydrogel formed by rat tail-derived type I collagen or, alternatively, by commercial Matrigel. The neural tissue is placed in the hydrogel with other brain tissue pieces or cell aggregates genetically modified to secrete a particular molecule that can generate a gradient inside the hydrogel. The present method is uncomplicated and generally reproducible, and only a few specific details need to be considered during its preparation. Moreover, the degree and behavior of axonal growth or neural migration can be observed directly using phase-contrast, fluorescence microscopy or immunocytochemical methods. This protocol can be carried out in 4 weeks.
本方案使用鼠尾来源的 I 型胶原水凝胶来分析发育神经生物学中的关键过程，如趋化性排斥和趋化性吸引。该方法基于在由鼠尾来源的 I 型胶原或商业 Matrigel 形成的 3D 水凝胶中培养来自胚胎或围生期早期的小鼠的小块脑组织。将神经组织与其他脑组织块或遗传修饰以分泌可在水凝胶内产生梯度的特定分子的细胞聚集体一起放置在水凝胶中。该方法简单且通常可重复，在制备过程中仅需考虑一些具体细节。此外，可以使用相差、荧光显微镜或免疫细胞化学方法直接观察轴突生长或神经迁移的程度和行为。本方案可在 4 周内完成。
