阵列型微拓扑结构基底的制备及其对神经细胞电生物物理特性的影响

摘要

UV lithography, silicon etching and soft lithography were adopted to fabricate micropillar arrayed cell culture substrates. With this approach, poly(dimethylsiloxane) (PDMS)topographic substrates with pillar featuring dimensions equal to or larger than 4 μm could be susccessfully fabricated, with a pillar height of 4 μm. The fabrication of substrates with a pillar feafuring dimension of 2 μm might represent the maximal fabrication capability of this method, as pillar lodging or missing could be found. With a simple tilting angle method, we fabricated a topographic substrate formed with a packed polystyrene microbead array. The arrayed beads could be as small as 2 μm in diameter or even in submicrometer range. Cell culture experiments showed that arrayed topographical substrates can pro mote adhesion or spreading, affecting resting membrane potential establishement and enhance the responsiveness of voltage-gated calcium channels of neuronal cells. The arrayed topographic substrates provide an efficient tool for engineering the microenvironments for sensor neuronal cells.%以紫外光光刻、硅蚀刻及软光刻技术制备了微柱阵列型细胞培养基底.实验发现,在4μm的结构高度下,当微柱特征尺寸大于或等于4 μm时,该法能制备结构规整清晰的聚二甲基硅氧烷微柱阵列型基底.特征尺寸为2μm的基底已经接近该法的极限加工能力,所加工的微柱阵列出现倒伏或缺失.采用一种简单的倾斜角法可以制备一种聚苯乙烯微球致密阵列型基底.该基底的微球直径可以小至2μm或亚微米尺度.细胞培养实验发现,拓扑结构基底能促进神经细胞粘附与铺展,影响细胞静息电位水平,增强电压依从式钙通道功能响应.阵列型拓扑结构基底为构建传感器细胞微环境提供了有效的工程化手段.