聚乳酸微小凹图式上人神经母细胞瘤细胞生长行为及电压依从式钙通道功能响应性
首发时间:2012-11-09
摘要:制备了聚乳酸(PLLA)准三维微小凹图式,研究了SH-SY5Y人神经母细胞瘤细胞与其复合后的生长特性、分化前后细胞形态和电压依从式钙通道(VGCCs)功能响应性。细胞的分化以1μM 9-顺式视黄酸进行诱导。以普通显微光学图片结合激光共聚焦技术研究细胞的形态铺展。以钙离子荧光染料钙绿-1和激光共聚焦显微技术评价SH-SY5Y细胞分化前后在微小凹图式和平面基底上VGCCs的功能响应性。实验结果表明,在分化前细胞在微小凹图式中呈二维(2D)、三维(3D)和近似二维(N2D)等三种生长方式。无论是分化前或后,微小凹图式内分布的细胞较之平面基底上的二维细胞表现出VGCCs功能响应性(反应幅度及阳性细胞比率)的明显降低。在分化前,无论是细胞铺展面积还是VGCCs功能响应性,N2D细胞的数值均介于2D细胞和3D细胞之间,提示细胞形态铺展和VGCCs响应性具有明显的相关性。本研究为基于神经细胞的生物传感器构建中细胞功能分化的工程化诱导提供了有效的途径。
关键词: 生物传感器 SH-SY5Y细胞 共聚焦显微镜 细胞铺展 电压依从式钙通道
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Growth characteristic and VGCC Responsiveness of Human Neuroblastoma Cells on Poly (l-lactic acid) Microwell Patterns
Abstract:In this paper,Quasi-three-dimensional poly-L-lactic acid (PLLA) microwell patterns were fabricated with lithography-based replica molding. SH-SY5Y human neuroblastoma cells were interfaced with the patterns and the cell growth characteristic, morphology and voltage-gated calcium channel (VGCC) responsiveness were evaluated both before and after differentiation. Cell differentiation was induced with 1μM 9-cis-retinoic acid. Cell spreading was analyzed in terms of projection areas under light microscopy in combination with laser scanning confocal microscopy. VGCC responsiveness was evaluated with confocal microscopy and a calcium fluorescent indicator, Calcium Green-1, both before and after differentiation. The results showed that, before differentiation, growth characteristics of SH-SY5Y cells on the microwell patterns could be categorized as two dimensional (2D), three dimensional (3D) and near two dimensional (N2D) according to the location of cells within the pattern. Cells within microwells showed a decreased VGCC responsiveness as compared to 2D cells on flat substrates, either before or after differentiation. For either projection areas or VGCC responsiveness, N2D cells consistently had an intermediate value between those for 2D and 3D cells, suggesting a correlative relation between cell morphology and VGCC responsiveness. This study provides an approach for engineering cell functionality in neural cell-based biosensor establishment.
Keywords: cell-based biosensor SH-SY5Y cells confocal microscopy cell spreading voltage-gated calcium channels
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聚乳酸微小凹图式上人神经母细胞瘤细胞生长行为及电压依从式钙通道功能响应性
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