Graduate School of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-0006, Japan.
J Biosci Bioeng. 2010 Aug;110(2):230-7. doi: 10.1016/j.jbiosc.2010.02.001. Epub 2010 Feb 26.
We describe the use of a microfluidic device to micropattern cells in a microchannel and investigated the behavior of these cells under a concentration gradient. The microfluidic device consisted of 3 parts: a branched channel for generating a stable concentration gradient, a main channel for culturing cells, and 2 side channels that flowed into the main channel. The main channel was coated with a cross-linked albumin that was initially cell-repellent but that could become cell-adherent by electrostatic adsorption of a polycation. A sheath flow stream, which was generated by introducing a polycation solution from the branched channel and a buffer solution from the 2 side channels, was used to change a specific region in the main channel from cell-repellent to cell-adhesive. In this way, cells attached to the central region along the main channel. The remaining surface was subsequently changed to cell-adhesive, thereby facilitating cell migration from a fixed location under a concentration gradient. We demonstrated that with this device, the gradient generator could be used to conduct simultaneous cytotoxic assays with anticancer agents; further, by combining this device with cell micropatterning, migration assays under a concentration gradient of biological factors could be conducted.
我们描述了一种使用微流控装置在微通道中对细胞进行微图案化的方法，并研究了这些细胞在浓度梯度下的行为。该微流控装置由 3 部分组成：用于产生稳定浓度梯度的分支通道、用于培养细胞的主通道和流入主通道的 2 个侧通道。主通道涂有交联白蛋白，最初对细胞具有排斥性，但通过聚阳离子的静电吸附可以变成细胞黏附性。鞘流由从分支通道引入的聚阳离子溶液和从 2 个侧通道引入的缓冲溶液产生，用于将主通道中的特定区域从细胞排斥性改变为细胞黏附性。这样，细胞就会附着在沿着主通道的中央区域。随后，剩余的表面被改变为细胞黏附性，从而促进细胞在浓度梯度下从固定位置迁移。我们证明，使用该装置，梯度发生器可用于同时进行抗癌药物的细胞毒性测定；此外，通过将该装置与细胞微图案化相结合，可以进行生物因子浓度梯度下的迁移测定。
