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以紫外光光刻、硅蚀刻及软光刻技术制备了聚乳酸(poly-L-lactide,PLLA)三维微小凹图式,考察人神经母细胞瘤细胞(SH-SY5Y)在微小凹结构上的生长与分布行为。以共聚焦显微技术结合钙离子荧光染料Calcium Green-1,AM评价SH-SY5Y细胞在PLLA三维微小凹图式和平面基底上电压依从式钙离子通道(VGCCs)的功能响应性。实验发现,分布在微小凹顶部的细胞为二维生长方式,凹内侧壁的为三维生长方式,凹内底部的为边界二维生长方式;以50 mmol/L高钾溶液去极化刺激发现,三维及边界二维细胞VGCCs阳性响应比率为75%和81%,较平面培养的91.2%明显降低。上述结果表明,PLLA三维微结构及细胞生长行为是影响SH-SY5Y细胞VGCCs功能响应性的重要因素,微小凹图式是模拟神经细胞三维生长环境进而构建三维细胞生物传感器的有效途径。
A poly-L-lactide (PLLA) three-dimensional micro-concave pattern was prepared by UV, Si and soft lithography. The effects of SH-SY5Y on the micro-concave structure Growth and distribution behavior. The functional responsiveness of SH-SY5Y cells to voltage-dependent calcium channels (VGCCs) on PLLA three-dimensional microgroove and planar substrates was evaluated by confocal microscopy and Calcium Green-1, AM. It was found that the cells distributed in the top of the micro concave were two-dimensional growth mode, the three-dimensional growth mode was in the concave inner side wall and the two-dimensional growth mode was in the bottom of the concave. With the depolarization stimulation of 50 mmol / L potassium solution, The positive response rates of VGCCs in three-dimensional and borderline two-dimensional cells were 75% and 81%, respectively, which were significantly lower than 91.2% in in-plane culture. The above results show that PLLA three-dimensional microstructure and cell growth behavior is an important factor affecting the functional responsiveness of VGCCs in SH-SY5Y cells. The micro-concave pattern is an effective way to simulate the three-dimensional growth environment of nerve cells and then to construct three-dimensional cell biosensors.