Coculture system for visualizing AD neurodegeneration process

开发了用于观察阿尔茨海默病神经退行性变过程的细胞培养底物。由PDMS制成的用于细胞培养的多个微室集成在带有64个嵌入微电极的衬底上。微室通过平行的PDMS微隧道相互连接。隧道结构高度设计为5um，以避免细胞体的迁移。取Wistar大鼠胚胎海马神经元，在基质上培养。神经细胞骨架特异性标志物- β 3-微管蛋白的免疫组织化学染色表明，神经突被成功引导进入微隧道并到达邻近的腔室。将Ab作用于其中一个微室，并通过细胞外记录和细胞内Ca瞬变来评估其活性。结果表明，钙离子浓度的过度升高可能是Ab引起的，这可能是神经变性的原因之一。

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.