Coculture system for visualizing AD neurodegeneration process

开发了用于可视化阿尔茨海默氏神经变性过程的产生和繁殖的细胞培养基质。将多个由PDMS制成的用于细胞培养的微室集成在具有64个嵌入式微电极的基底上。微室通过平行的PDMS微隧道彼此连接。隧道结构的高度设计为5 μ m，以避免细胞体的迁移。海马神经元取自Wistar大鼠胚胎并在基质上培养。β 3-微管蛋白是神经元细胞骨架的特异性标记物，其免疫组织化学染色表明，神经突被成功地引导到微隧道中并到达相邻的腔室。将Ab施加到其中一个微室中，并通过细胞外记录和细胞内Ca瞬变来评价活性。结果提示，抗坏血酸可引起钙离子浓度过度升高，这可能是导致神经元变性的原因之一。

Cell-culture substrates for visualizing generation and propagation of Alzheimer's neuro-degeneration processes were developed. Multiple micro-chambers made of PDMS for cell culture were integrated on a substrate with 64 embedded micro-electrodes. The micro-chambers were connected each other via parallel PDMS micro-tunnels. The height of the tunnel structure was designed as 5 um to avoid migration of cell bodies. Hippocampal neurons were taken from Wistar rat embryos and cultured on the substrates. Immuno-histochemical staining with beta3-tubulin, which was a specific marker for neuronal cytoskeleton, suggested that neurites were successfully guided into the micro-tunnels and reached the neighbor chamber. Ab was applied to one of the micro-chambers and the activity was evaluated by extracellular recording and intracellular Ca transients. The results suggested that excess elevation of Ca concentration was induced by Ab application, which might be one of the causes of neuro-degeneration.