Harrill Joshua A, Mundy William R
Systems Biology Branch, Integrated Systems Toxicology Division, National Health and Environmental Effects Research Laboratories, United States Environmental Protection Agency, Research Triangle Park, NC, USA.
Methods Mol Biol. 2011;758:331-48. doi: 10.1007/978-1-61779-170-3_23.
In vitro test methods can provide a rapid approach for the screening of large numbers of chemicals for their potential to produce toxicity. In order to identify potential developmental neurotoxicants, assessment of critical neurodevelopmental processes, such as neuronal differentiation and growth has been proposed. PC12 cells have been widely used to study the neurotrophic factor-induced signaling pathways that control differentiation, and as in vitro models to detect the effect of chemicals on neurite outgrowth. Upon exposure to nerve growth factor (NGF), PC12 cells cease to proliferate, extend multiple neurites, and acquire the properties of sympathetic neurons. Measurement of the number and length of neurites during exposure to NGF provides a quantitative assessment of neuronal differentiation and growth. Differentiation and neurite outgrowth can be measured using simple contrast microscopy in live cells, or using automated imaging systems in cells prepared with immunocytochemistry.
体外测试方法可为大量化学物质产生毒性的潜力筛选提供一种快速途径。为了识别潜在的发育性神经毒物，有人提出评估关键的神经发育过程，如神经元分化和生长。PC12细胞已被广泛用于研究控制分化的神经营养因子诱导的信号通路，并作为体外模型来检测化学物质对神经突生长的影响。暴露于神经生长因子（NGF）后，PC12细胞停止增殖，长出多个神经突，并获得交感神经元的特性。在暴露于NGF期间测量神经突的数量和长度可对神经元分化和生长进行定量评估。可以使用活细胞中的简单相差显微镜，或使用免疫细胞化学制备的细胞中的自动成像系统来测量分化和神经突生长。
