Phillips James B
Department of Biomaterials and Tissue Engineering, UCL Eastman Dental Institute, University College London, 256 Gray's Inn Road, London, WC1X 8LD, UK,
Methods Mol Biol. 2014;1162:113-24. doi: 10.1007/978-1-4939-0777-9_9.
Methods are described for the generation and analysis of 3D co-culture models in which astrocyte and neuronal behavior can be studied. Cells may be obtained from a variety of sources, then cultured within collagen hydrogels to explore cellular responses and interactions in response to substances under test or under conditions that mimic physiological or pathological environments. Cell populations are labelled then either mixed within gels or arranged as separate adjacent populations, with further options including directing the self-alignment of cells to form anisotropic 3D cultures. Immunofluorescence staining and confocal microscopy can be used to capture image data from 3D structures and detailed protocols are provided for obtaining reliable results. Finally, 3D image analysis of confocal microscopy data is discussed, providing guidance on how astrocyte and neuronal features can be quantified.
本文描述了用于生成和分析三维共培养模型的方法，在该模型中可以研究星形胶质细胞和神经元的行为。细胞可以从多种来源获得，然后在胶原蛋白水凝胶中培养，以探索细胞对受试物质或模拟生理或病理环境条件的反应及相互作用。对细胞群体进行标记，然后将其混合在凝胶中或排列成相邻的独立群体，还有进一步的选项，包括引导细胞自我排列以形成各向异性的三维培养物。免疫荧光染色和共聚焦显微镜可用于从三维结构中获取图像数据，并提供了详细的方案以获得可靠的结果。最后，讨论了共聚焦显微镜数据的三维图像分析，为如何量化星形胶质细胞和神经元特征提供指导。
