A microchip-based system for immobilizing PC 12 cells and amperometrically detecting catecholamines released after stimulation with calcium
A microchip-based system for immobilizing PC 12 cells and amperometrically detecting catecholamines released after stimulation with calcium
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DOI：
10.1002/elan.200403231
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发表时间：
2005-07-01
期刊：
影响因子：
3
通讯作者：
Martin, RS
中科院分区：
文献类型：
--
作者：
Li, MW;Spence, DM;Martin, RS
在本文中，我们描述了一种基于微芯片的系统，用于电流监测的量的儿茶酚胺释放大鼠嗜铬细胞瘤（PC 12）细胞。该系统的关键是一种新的，但简单的方法固定PC 12细胞在聚二甲基硅氧烷（PDMS）为基础的微通道。该过程包括用胶原蛋白选择性地涂覆微通道，然后在PDMS结构上引入PC 12细胞，其中细胞仅固定在通道的涂覆部分上。然后可以将细胞涂覆的微通道可逆地密封到含有用于安培检测的电极的玻璃板上，从而得到具有集成微电极的固定化细胞反应器。用碳墨微成型的Nafion涂层微电极测量钙诱导的细胞释放的儿茶酚。当用钙溶液刺激细胞时，固定在微通道中的不同浓度的PC 12细胞会释放20至160 μ M的儿茶酚胺。这种微芯片方法导致了一种三维培养，可以与这种或其他细胞系一起使用，以研究外部刺激对神经递质释放的影响。
In this paper, we describe a microchip-based system for amperometrically monitoring the amount of catecholamines released from rat pheochromocytoma (PC 12) cells. Key to this system is a novel, yet simple method for the immobilization of PC 12 cells in poly(dimethylsiloxane) (PDMS)-based microchannels. The procedure involves selectively coating microchannels with collagen followed by introduction of PC 12 cells over the PDMS structure, with the cells being immobilized only on the coated portion of the channels. The cell-coated microchannels can then be reversibly sealed to a glass plate containing electrodes for amperometric detection, resulting in an immobilized cell reactor with integrated microelectrodes. Nafion-coated microelectrodes made by micromolding of carbon inks were used to measure calcium-induced catecholarnine release from the cells. Varying concentrations of PC 12 cells immobilized in the microchannels led to a catecholamine release ranging from 20 to 160 mu M when the cells were stimulated with a calcium solution. This microchip approach leads to a three-dimensional culture that can be used with this or other cells lines to study the effect of external stimuli on neurotransmitter release.